Use of short chain fatty acids for the treatment and prevention of diseases and disorders

ABSTRACT

The invention relates to compositions and methods that include short chain fatty acids (SCFAs), related therapeutic compounds, and other compounds, including combinations thereof, for the treatment or prevention of diseases or disorders.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of priority under 35 U.S.C. 365(c)to International Patent Application No. PCT/US2018/015383, filed on Jan.26, 2018, which is entitled to priority under 35 U.S.C. § 119(e) to U.S.Provisional Patent Application No. 62/451,192 filed Jan. 27, 2017, U.S.Provisional Patent Application No. 62/510,867 filed May 25, 2017, U.S.Provisional Patent Application No. 62/510,872 filed May 25, 2017, U.S.Provisional Patent Application No. 62/530,371 filed Jul. 10, 2017, U.S.Provisional Patent Application No. 62/539,572 filed Aug. 1, 2017, andU.S. Provisional Patent Application No. 62/588,961 filed Nov. 21, 2017,the disclosures of which are incorporated herein by reference in theirentirety.

BACKGROUND OF THE INVENTION

Short chain fatty acids (SCFAs) are the main metabolic products ofanaerobic bacteria fermentation in the intestine, and have been shown tomodulate different processes (such as metabolic functions andhomeostasis) in the gastrointestinal (GI) tract in adipose tissue,immune and nervous systems. Quantitative and qualitative changes in thegut microbiome and, consequently, changes in the concentrations of theproduced metabolites have been suggested to promote the development ofpathological conditions including inflammatory bowel disease (IBD),colon cancer, obesity and type 1 and 2 diabetes mellitus.

SCFAs are saturated aliphatic acids consisting of one polar carboxylicacid moiety and hydrophobic hydrocarbon chain among which acetate (C2),propionate (C3) and butyrate (C4) are the most common and well-studiedmolecules.

The SCFAs affect several cellular processes including gene expression,chemotaxis, differentiation, proliferation and apoptosis via activationof G protein coupled receptors (GPCRs), inhibition of histonedeacetylases (HDACs) and by binding to the butyrate-responsive elementsin the gene promoter regions of some transcriptional factors (which mayexplain the pleiotropic effects of butyrate). SCFAs are importantregulators of inflammation by controlling migration of immune cellstoward inflammatory sites as well as modulating their activation state.Importantly, SCFAs influence the balance between pro- andanti-inflammatory cells and serve as a means of communication betweenmicrobiota and the immune system. The principle mechanisms through whichSCFAs exert anti-inflammatory effects are suppression of TNFα and NF-κBactivation, the inhibition of IFN-γ production, and upregulation of theperoxisome proliferator-activated receptor γ (PPARγ, a nuclear hormonereceptor capable to inhibit NF-κB dependent transcriptional activation.)

Current SCFA therapeutic compounds have limited or inadequate efficacybecause of the rapid absorption in the small intestine and a very shorthalf-life. Thus there is a need in the art for safe effectivecompositions comprising SCFA for use in the treatment or prevention ofdiseases and disorders. The present invention addresses this unmet need.

SUMMARY OF THE INVENTION

In one embodiment, the invention relates to a composition comprising atleast one compound selected from the group consisting of: a short chainfatty acid (SCFA), a SCFA precursor, a SCFA biosynthesis precursor, acompound comprising a SFCA moiety, a derivative thereof, and acombination thereof.

In one embodiment, the composition comprises at least one SCFA moietylinked to at least one additional moiety.

In one embodiment, the at least one additional moiety comprisespolyethylene glycol (PEG).

In one embodiment, the composition hydrolyzes under a low pH conditionto yield PEG and a SCFA.

In one embodiment, the SCFA or SCFA moiety comprises at least oneselected from the group consisting of: acetic acid, butyric acid (BA),C3-C12 fatty acids, C3-C10 fatty acids, C3-C8 fatty acids, methoxyaceticacid, valproic acid (VPA), propionic acid, 3-methoxypropionic acid,ethoxyacetic acid, formic acid, isobutyric acid, tributyrin, butyrate,propionate, N-acetylbutyrate (as well as other forms of butyrate, e.g.,phenylbutyrate, isobutyrate, pivaloyloxymethyl butyrate, monoacetoneglucose 3-butyrate), isovaleric acid, valeric acid, isocaproic acid,caproic acid, lactic acid, succinic acid, pyruvic acid, octanoic acid,dodecanoic acid, (4R)-4-hydroxypentanoic acid, 2-ethylhydracrylic acid,2-hydroxy-3-methylpentanoate, 2-hydroxy-3-methylpentanoic acid,2-methylbut-2-enoic acid, 2-oxobutanoic acid, 3-hydroxypentanoic acid,3-methylbut-2-enoic acid, butenoic acid, methylbutyric acid,dimethylbutyric acid, pentadienoic acid, pentenoic acid, pivalic acid,propynoic acid and a combination thereof.

In one embodiment, the SCFA precursor or SCFA precursor moiety of theSCFA precursor derivative comprises at least one selected from the groupconsisting of: a salt of lactate, a salt of succinate, a salt offormate, 1,2-propenedol, trypamine, indole, indole-3-acetate, and acombination thereof.

In one embodiment, the SCFA biosynthesis precursor or SCFA biosynthesisprecursor moiety of the SCFA biosynthesis precursor derivative comprisesan acetyl-CoA carboxylase inhibitor, an adenosine monophosphate kinase(AMPK) activator, vitamin D, or a combination thereof.

In one embodiment, the composition comprises butyrate.

In one embodiment, the composition comprises at least two SCFAs.

In one embodiment, the composition comprises butyrate and propionate.

In one embodiment, the invention relates to a pharmaceutical compositioncomprising at least one compound selected from: a short chain fatty acid(SCFA), a SCFA precursor, a SCFA biosynthesis precursor, a compoundcomprising a SFCA moiety, a derivative thereof, and a combinationthereof.

In one embodiment, the pharmaceutical composition comprises at least oneSCFA moiety linked to at least one additional moiety. In one embodiment,the at least one additional moiety comprises polyethylene glycol (PEG).

In one embodiment, the pharmaceutical composition hydrolyzes under a lowpH condition to yield PEG and a SCFA.

In one embodiment, the SCFA or SCFA moiety comprises at least oneselected from the group consisting of: acetic acid, butyric acid (BA),C3-C12 fatty acids, C3-C10 fatty acids, C3-C8 fatty acids, methoxyaceticacid, valproic acid (VPA), propionic acid, 3-methoxypropionic acid,ethoxyacetic acid, formic acid, isobutyric acid, tributyrin, butyrate,propionate, N-acetylbutyrate (as well as other forms of butyrate, e.g.,phenylbutyrate, isobutyrate, pivaloyloxymethyl butyrate, monoacetoneglucose 3-butyrate), isovaleric acid, valeric acid, isocaproic acid,caproic acid, lactic acid, succinic acid, pyruvic acid, octanoic acid,dodecanoic acid, (4R)-4-hydroxypentanoic acid, 2-ethylhydracrylic acid,2-hydroxy-3-methylpentanoate, 2-hydroxy-3-methylpentanoic acid,2-methylbut-2-enoic acid, 2-oxobutanoic acid, 3-hydroxypentanoic acid,3-methylbut-2-enoic acid, butenoic acid, methylbutyric acid,dimethylbutyric acid, pentadienoic acid, pentenoic acid, pivalic acid,propynoic acid and a combination thereof.

In one embodiment, the SCFA precursor or SCFA precursor moiety of theSCFA precursor derivative comprises at least one selected from the groupconsisting of: a salt of lactate, a salt of succinate, a salt offormate, 1,2-propenedol, trypamine, indole, indole-3-acetate, and acombination thereof.

In one embodiment, the SCFA biosynthesis precursor or SCFA biosynthesisprecursor moiety of the SCFA biosynthesis precursor derivative comprisesan acetyl-CoA carboxylase inhibitor, an adenosine monophosphate kinase(AMPK) activator, vitamin D, or a combination thereof.

In one embodiment, the pharmaceutical composition comprises 100milligrams (mg) to 6 grams (g) of at least one SCFA.

In one embodiment, the pharmaceutical composition comprises butyrate. Inone embodiment, the pharmaceutical composition comprises 900 to 1800 mgbutyrate.

In one embodiment, the pharmaceutical composition comprises at least twoSCFAs.

In one embodiment, the pharmaceutical composition comprises butyrate andpropionate. In one embodiment, the pharmaceutical composition comprises900 to 1800 mg butyrate and 100 mg to 200 mg propionate.

In one embodiment, the pharmaceutical composition further comprises apharmaceutically acceptable excipient.

In one embodiment, the pharmaceutical composition is an enteric coated,extended release capsule.

In one embodiment, the pharmaceutical composition comprises 900 mgbutyrate, 100 mg propionate, 10 mg apremilast, 10 mg magnesium, and 50IU vitamin D3.

In one embodiment, the pharmaceutical composition is formulated as aneyedrop. In one embodiment, the eyedrop comprises 10 micromolar (μM) to100 μM butyrate. In one embodiment, the eyedrop comprises 10 μM to 100μM propionate. In one embodiment, the eyedrop comprises 10 μM to 100 μMbutyrate and 10 to 100 μM propionate.

In one embodiment, the invention relates to a method for treating orpreventing a disease or disorder, comprising administering to a subjecta composition or pharmaceutical composition comprising at least onecompound selected from: a short chain fatty acid (SCFA), a SCFAprecursor, a SCFA biosynthesis precursor, a compound comprising a SFCAmoiety, a derivative thereof, and a combination thereof.

In one embodiment, the disease or disorder is a skin disease ordisorder, an allergic or autoimmune disease or disorder, an eye diseaseor disorder, an adverse effect associated with immunotherapy, cancer ora combination thereof.

In one embodiment, the method comprises administering a composition orpharmaceutical composition comprising 100 mg to 6 g butyrate.

In one embodiment, the method comprises administering the composition orpharmaceutical composition daily for at least one week.

In one embodiment, the method comprises administering the composition orpharmaceutical composition one to three times daily for at least oneweek.

In one embodiment, the method comprises further comprising administeringat least one additional treatment or therapeutic agent to the subject.

In one embodiment, the at least one additional treatment isimmunotherapy.

In one embodiment, the invention relates to a method for treating orpreventing a skin disease or disorder, comprising administering acomposition comprising at least 900 mg of butyrate and at least 100 mgpropionate in enteric coated, extended release capsules, 3 times/day forat least 2 weeks. In one embodiment, the method further comprisesadministering a dose of at least 900 mg of butyrate, twice a day for atleast 2 weeks, following the completion of the first dosage regimen.

In one embodiment, the invention relates to a method for treating orpreventing a skin disease or disorder, comprising administering a doseof at least 900 mg of butyrate, at least 100 mg propionate, and at least10 mg apremilast in enteric coated, extended release capsules, 3times/day for at least 2 weeks.

In one embodiment, the invention relates to a method for treating orpreventing an eye disease or disorder comprising: administering a dailyoral dose of at least 4 g of butyrate and at least 1.5 g propionate inenteric coated, extended release capsules, 3 times/day for at least 2weeks.

In one embodiment, the invention relates to a method for treating orpreventing an eye disease or disorder comprising administering eyedropscontaining at least 20 μM of butyrate and at least 10 μM propionate atleast 2 times/day for at least 3 days.

In one embodiment, the invention relates to a method for treating orpreventing an adverse effect associated with immunotherapy comprisingadministering a daily oral dose of at least 5 g of butyrate and at least2 g propionate in enteric coated, extended release capsules, 3 times/dayfor at least 2 weeks. In one embodiment, the administering is performedprior to an immunotherapy, concurrent with an immunotherapy, subsequentto an immunotherapy, or a combination thereof.

In one embodiment, the invention relates to a method for treating orpreventing an allergic disease, an autoimmune disease or asthmacomprising: administering a daily oral dose of at least 1 of butyrate,at least 0.5 g propionate, and at least 0.5 g acetate in enteric coated,extended release capsules, 3 times/day for at least 2 weeks.

In one embodiment, the invention relates to a method for treating orpreventing an allergic disease, an autoimmune disease and asthma in aC-section delivered neonate comprising: administering to the neonate adaily oral dose of at least 80 mg of butyrate, at least 20 mgpropionate, and at least 20 mg acetate as a nutraceutical.

In one embodiment, the invention relates to a method for treating orpreventing vasculitis comprising administering a daily oral dose of atleast 5 g of butyrate in enteric coated, extended release capsules, 3times/day for at least 2 weeks. In one embodiment, the method furthercomprises administering a daily oral dose of at least 3 g of butyrate,twice a day for at least 2 weeks, following the completion of the firstdosage regimen.

BRIEF DESCRIPTION OF THE DRAWINGS

The following detailed description of embodiments of the invention willbe better understood when read in conjunction with the appendeddrawings. It should be understood that the invention is not limited tothe precise arrangements and instrumentalities of the embodiments shownin the drawings.

FIG. 1 depicts a diagram showing signaling pathways that are affected bySCFAs. Of the proteins included on the diagram, IL-18, TLR3, IFN-γ,TNFα, TGF-β, MyD88, PI3K/Akt, JAK/STAT, Smad 2/3, Smad 4 and IL-10 werefound to be downregulated by SCFA treatment. The levels of IL-6, TRIF,PKR, TRAF2, TAK1 and TRAF6 were not evaluated for this study.

FIG. 2, comprising FIG. 2A through FIG. 2C, depicts images from apsoriasis patient undergoing a butyrate treatment regimen. The treatmentregimen used was one (1) 600 mg butyrate pill 3 times daily for 16 days.FIG. 2A depicts images of the patient's left elbow before and aftertreatment. FIG. 2B depicts images of the patient's right elbow beforeand after treatment. FIG. 2C depicts an image of the re-emergence ofpsoriasis on the patient's left elbow after 20 days the patient ceasedthe butyrate treatment regimen.

FIG. 3 depicts images from another psoriasis patient undergoing abutyrate treatment regimen. The treatment regimen used was two (2) 600mg butyrate pill 3 times daily for 10 days, then regimen was one (1)butyrate pill, three times/day for three weeks.

FIG. 4 depicts images from a third psoriasis patient undergoing abutyrate treatment regimen. The treatment regimen used was two (2) 600mg butyrate pills 3 times daily for 7 days then one (1) 600 mg butyratepill 3 times daily for 24 days. Images are of the patient's left elbowbefore and after treatment. The patient consumed elevated doses ofalcohol during the treatment period.

FIG. 5, comprising FIG. 5A through FIG. 5C, depicts images from a fourthpatient with severe psoriasis over about 50% of the skin undergoing abutyrate treatment regimen. Patient started regimen as two (2) 600 mgbutyrate pills 3 times daily for 4 weeks followed by one (1) 600 mgbutyrate pill 3 times daily for several months. There was significantimprovement, and no side effects were noted during the treatment forlong period of time. To confirm that observed positive effects were dueto treatment with our formulation, patient was asked to stop regimen.Psoriasis came back in about 20-25 days (pictures before). Patientstarted regimen as two (2) 600 mg butyrate pills 3 times daily for 10days then one (1) 600 mg butyrate pills 3 times daily for 3 weeks andpsoriasis disappeared (pictures after). FIG. 5A depicts images of thepatient's left hip before and after treatment the second treatmentregimen. FIG. 5B depicts images of the patient's right hip before andafter the second treatment regimen. FIG. 5C depicts images of thepatient's tailbone before and after the second treatment regimen.

FIG. 6 depicts images from a patient with a plaque psoriasis. An initialtreatment regimen used was two (2) 600 mg butyrate pills 3 times dailyfor 20 days, then regimen was with a reduced dose, one (1) 600 mgbutyrate pills 3 times daily for 25 days. Significant improvement wasobserved 3-5 months after he stopped his treatment. Patient was nottaking any medicine before (at least 3 months), during and aftertreatment (at least 2 months).

DETAILED DESCRIPTION

In one embodiment, the invention relates to compositions comprising atleast one SCFA for the treatment or prevention of one or more diseasesand/or disorders in a subject.

In one embodiment, the SCFA comprises at least one selected from thegroup including, but not limited to: a SCFA, a SCFA precursor, a SCFAbiosynthesis precursor, a compound comprising a SCFA moiety, aderivative thereof, a salt thereof, an ester thereof, a conjugate basethereof (for example, PEGylated conjugates), and a combination thereof.In one embodiment, a SCFA or SCFA moiety includes, but is not limitedto, acetic acid, butyric acid (BA), C3-C12 fatty acids, C3-C10 fattyacids, C3-C8 fatty acids, methoxyacetic acid, valproic acid (VPA),propionic acid, 3-methoxypropionic acid, ethoxyacetic acid, formic acid,isobutyric acid, tributyrin, butyrate, propionate, N-acetylbutyrate (aswell as other forms of butyrate, e.g., phenylbutyrate, isobutyrate,pivaloyloxymethyl butyrate, monoacetone glucose 3-butyrate), isovalericacid, valeric acid, isocaproic acid, caproic acid, lactic acid, succinicacid, pyruvic acid, octanoic acid, dodecanoic acid,(4R)-4-hydroxypentanoic acid, 2-ethylhydracrylic acid,2-hydroxy-3-methylpentanoate, 2-hydroxy-3-methylpentanoic acid,2-methylbut-2-enoic acid, 2-oxobutanoic acid, 3-hydroxypentanoic acid,3-methylbut-2-enoic acid, butenoic acid, methylbutyric acid,dimethylbutyric acid, pentadienoic acid, pentenoic acid, pivalic acid,propynoic acid and a combination thereof. In one embodiment, the SCFA orSCFA moiety includes compounds or structures with at least 12 carbonatoms, at least 11 carbon atoms, at least 10 carbon atoms, at least 9carbon atoms, at least 8 carbon atoms, at least 7 carbon atoms, at least6 carbon atoms, at least 5 carbon atoms, at least 4 carbon atoms, atleast 3 carbon atoms, and at least 2 carbon atoms. In one embodiment,the SCFA or SCFA moiety is not a branched fatty acid. In one embodiment,the SCFA or SCFA moiety includes compounds or structures with less than13 carbon atoms, less than 12 carbon atoms, less than 11 carbon atoms,less than 10 carbon atoms, less than 9 carbon atoms, less than 8 carbonatoms, or less than 7 carbon atoms. In one embodiment, the SCFA or SCFAmoiety is not a branched fatty acid. In one embodiment, the SCFA or SCFAmoiety is a branched fatty acid.

In one embodiment, the invention relates to methods for the treatment orprevention of one or more diseases and/or disorders in a subject,comprising administering to a subject at least one compositioncomprising a SCFA. In one embodiment, at least one compositioncomprising a SCFA is administered to a subject in combination with atleast one additional agent or therapy, for the treatment or preventionof a disease and/or disorder in a subject. In one embodiment, at leastone SCFA and at least one additional therapeutic agent are administeredtogether in one composition. In one embodiment, at least one SCFA and atleast one additional therapeutic agent are administered separately, astwo or more compositions.

In one embodiment, a subject is a human. In one embodiment, a subject isa non-human animal.

Definitions

It is to be understood that the figures and descriptions of the presentinvention have been simplified to illustrate elements that are relevantfor a clear understanding of the present invention, while eliminating,for the purpose of clarity, many other elements found in the invention.Those of ordinary skill in the art may recognize that other elementsand/or steps are desirable and/or required in implementing the presentinvention. However, because such elements and steps are well known inthe art, and because they do not facilitate a better understanding ofthe present invention, a discussion of such elements and steps is notprovided herein. The disclosure herein is directed to all suchvariations and modifications to such elements and methods known to thoseskilled in the art.

Unless defined elsewhere, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, exemplary methods andmaterials are described.

As used herein, each of the following terms has the meaning associatedwith it in this section.

The articles “a” and “an” are used herein to refer to one or to morethan one (i.e., to at least one) of the grammatical object of thearticle. By way of example, “an element” means one element or more thanone element.

“About” as used herein when referring to a measurable value such as anamount, a temporal duration, and the like, is meant to encompassvariations of ±20%, ±10%, ±5%, ±1%, or ±0.1% from the specified value,as such variations are appropriate.

The term “abnormal” when used in the context of organisms, tissues,cells or components thereof, refers to those organisms, tissues, cellsor components thereof that differ in at least one observable ordetectable characteristic (e.g., age, treatment, time of day, etc.) fromthose organisms, tissues, cells or components thereof that display the“normal” (expected) respective characteristic. Characteristics which arenormal or expected for one cell or tissue type, might be abnormal for adifferent cell or tissue type.

The term “adverse effect of cancer immunotherapy,” as used herein, mayrefer to at least one of the following types of adverse effect of cancerimmunotherapy: cytokine release syndrome (CRS), neurological toxicity,on-target/off-tumor recognition, anaphylaxis, graft versus host disease(GVHD), off-target antigen recognition, and macrophage activationsyndrome (MAS).

By “allergy” or “allergic disease or disorder” is meant one or more of anumber of conditions caused by hypersensitivity of the immune system toone or more substances in the environment.

A subject is “at risk for” developing a condition if there is anincreased probability that the individual will develop the conditioncompared to a population (e.g., the general population, an age-matchedpopulation, a population of the same sex). The increased probability canbe due to one or a combination of factors including the presence ofspecific alleles/mutations of a gene or exposure to a particularenvironment.

By an “autoimmune disease or disorder” is meant an immune responseagainst a self-antigen that results in inflammation or destruction ofhealthy tissue in a subject. Desirably the subject is a mammal, such asa human. Exemplary autoimmune diseases include, but are not limited to,arthritis (e.g., rheumatoid arthritis such as acute arthritis, chronicrheumatoid arthritis, gouty arthritis, acute gouty arthritis, chronicinflammatory arthritis, degenerative arthritis, infectious arthritis,Lyme arthritis, proliferative arthritis, psoriatic arthritis, vertebralarthritis, and juvenile-onset rheumatoid arthritis, osteoarthritis,arthritis chronica progrediente, arthritis deformans, polyarthritischronica primaria, reactive arthritis, and ankylosing spondylitis),inflammatory hyperproliferative skin diseases, psoriasis such as plaquepsoriasis, gutatte psoriasis, pustular psoriasis, psoriasis vulgaris,inverse psoriasis, erythrodermic psoriasis, seborrheic psoriasis andpsoriasis of the nails, dermatitis including contact dermatitis, chroniccontact dermatitis, allergic dermatitis, allergic contact dermatitis,dermatitis herpetiformis, and atopic dermatitis, x-linked hyper IgMsyndrome, urticaria such as chronic allergic urticaria and chronicidiopathic urticaria, including chronic autoimmune urticaria,polymyositis/dermatomyositis, juvenile dermatomyositis, toxic epidermalnecrolysis, scleroderma (including systemic scleroderma), sclerosis suchas systemic sclerosis, multiple sclerosis (MS) such as spino-optical MS,primary progressive MS (PPMS), and relapsing remitting MS (RRMS),progressive systemic sclerosis, sclerosis disseminata, and ataxicsclerosis, inflammatory bowel disease (IBD) (for example, Crohn'sdisease, autoimmune-mediated gastrointestinal diseases, colitis such asulcerative colitis, colitis ulcerosa, microscopic colitis, collagenouscolitis, colitis polyposa, necrotizing enterocolitis, and transmuralcolitis, and autoimmune inflammatory bowel disease), pyodermagangrenosum, erythema nodosum, primary sclerosing cholangitis,episcleritis, respiratory distress syndrome, including adult or acuterespiratory distress syndrome (ARDS), meningitis, inflammation of all orpart of the uvea, iritis, choroiditis, an autoimmune hematologicaldisorder, rheumatoid spondylitis, sudden hearing loss, IgE-mediateddiseases such as anaphylaxis and allergic and atopic rhinitis,encephalitis such as Rasmussen's encephalitis and limbic and/orbrainstem encephalitis, uveitis, such as anterior uveitis, acuteanterior uveitis, granulomatous uveitis, nongranulomatous uveitis,phacoantigenic uveitis, posterior uveitis, or autoimmune uveitis,glomerulonephritis (GN) with and without nephrotic syndrome such aschronic or acute glomerulonephritis such as primary GN, immune-mediatedGN, membranous GN (membranous nephropathy), idiopathic membranous GN oridiopathic membranous nephropathy, membrano- or membranous proliferativeGN (MPGN), including Type I and Type II, and rapidly progressive GN,allergic conditions, allergic reaction, eczema including allergic oratopic eczema, asthma such as asthma bronchiale, bronchial asthma, andauto-immune asthma, conditions involving infiltration of T cells andchronic inflammatory responses, chronic pulmonary inflammatory disease,autoimmune myocarditis, leukocyte adhesion deficiency, systemic lupuserythematosus (SLE) or systemic lupus erythematodes such as cutaneousSLE, subacute cutaneous lupus erythematosus, neonatal lupus syndrome(NLE), lupus erythematosus disseminatus, lupus (including nephritis,cerebritis, pediatric, non-renal, extra-renal, discoid, alopecia),juvenile onset (Type I) diabetes mellitus, including pediatricinsulin-dependent diabetes mellitus (IDDM), adult onset diabetesmellitus (Type II diabetes), autoimmune diabetes, idiopathic diabetesinsipidus, immune responses associated with acute and delayedhypersensitivity mediated by cytokines and T-lymphocytes, tuberculosis,sarcoidosis, granulomatosis including lymphomatoid granulomatosis,Wegener's granulomatosis, agranulocytosis, vasculitides, includingvasculitis (including large vessel vasculitis (including polymyalgiarheumatica and giant cell (Takayasu's) arteritis), medium vesselvasculitis (including Kawasaki's disease and polyarteritis nodosa),microscopic polyarteritis, CNS vasculitis, necrotizing, cutaneous, orhypersensitivity vasculitis, systemic necrotizing vasculitis, andANCA-associated vasculitis, such as Churg-Strauss vasculitis or syndrome(CSS)), temporal arteritis, aplastic anemia, autoimmune aplastic anemia,Coombs positive anemia, Diamond Blackfan anemia, hemolytic anemia orimmune hemolytic anemia including autoimmune hemolytic anemia (AIHA),pernicious anemia (anemia perniciosa), Addison's disease, pure red cellanemia or aplasia (PRCA), Factor VIII deficiency, hemophilia A,autoimmune neutropenia, pancytopenia, leukopenia, diseases involvingleukocyte diapedesis, CNS inflammatory disorders, multiple organ injurysyndrome such as those secondary to septicemia, trauma or hemorrhage,antigen-antibody complex-mediated diseases, anti-glomerular basementmembrane disease, anti-phospholipid antibody syndrome, allergicneuritis, Bechet's or Behcet's disease, Castleman's syndrome,Goodpasture's syndrome, Reynaud's syndrome, Sjogren's syndrome,Stevens-Johnson syndrome, pemphigoid such as pemphigoid bullous and skinpemphigoid, pemphigus (including pemphigus vulgaris, pemphigusfoliaceus, pemphigus mucus-membrane pemphigoid, and pemphiguserythematosus), autoimmune polyendocrinopathies, Reiter's disease orsyndrome, immune complex nephritis, antibody-mediated nephritis,neuromyelitis optica, polyneuropathies, chronic neuropathy such as IgMpolyneuropathies or IgM-mediated neuropathy, thrombocytopenia (asdeveloped by myocardial infarction patients, for example), includingthrombotic thrombocytopenic purpura (TTP) and autoimmune orimmune-mediated thrombocytopenia such as idiopathic thrombocytopenicpurpura (ITP) including chronic or acute ITP, autoimmune disease of thetestis and ovary including autoimmune orchitis and oophoritis, primaryhypothyroidism, hypoparathyroidism, autoimmune endocrine diseasesincluding thyroiditis such as autoimmune thyroiditis, Hashimoto'sdisease, chronic thyroiditis (Hashimoto's thyroiditis), or subacutethyroiditis, autoimmune thyroid disease, idiopathic hypothyroidism,Grave's disease, polyglandular syndromes such as autoimmunepolyglandular syndromes (or polyglandular endocrinopathy syndromes),paraneoplastic syndromes, including neurologic paraneoplastic syndromessuch as Lambert-Eaton myasthenic syndrome or Eaton-Lambert syndrome,stiff-man or stiff-person syndrome, encephalomyelitis such as allergicencephalomyelitis or encephalomyelitis allergica and experimentalallergic encephalomyelitis (EAE), myasthenia gravis such asthymoma-associated myasthenia gravis, cerebellar degeneration,neuromyotonia, opsoclonus or opsoclonus myoclonus syndrome (OMS), andsensory neuropathy, multifocal motor neuropathy, Sheehan's syndrome,lymphoid interstitial pneumonitis, bronchiolitis obliterans(non-transplant) vs NSIP, Guillain-Barre syndrome, Berger's disease (IgAnephropathy), idiopathic IgA nephropathy, linear IgA dermatosis, primarybiliary cirrhosis, pneumonocirrhosis, autoimmune enteropathy syndrome,Celiac disease, Coeliac disease, celiac sprue (gluten enteropathy),refractory sprue, idiopathic sprue, cryoglobulinemia, amylotrophiclateral sclerosis (ALS; Lou Gehrig's disease), coronary artery disease,autoimmune ear disease such as autoimmune inner ear disease (AIED),autoimmune hearing loss, opsoclonus myoclonus syndrome (OMS),polychondritis such as refractory or relapsed polychondritis, pulmonaryalveolar proteinosis, amyloidosis, scleritis, a non-cancerouslymphocytosis, a primary lymphocytosis, which includes monoclonal B celllymphocytosis (e.g., benign monoclonal gammopathy and monoclonalgarnmopathy of undetermined significance, MGUS), peripheral neuropathy,paraneoplastic syndrome, channelopathies such as epilepsy, migraine,arrhythmia, muscular disorders, deafness, blindness, periodic paralysis,and channelopathies of the CNS, autism, inflammatory myopathy, focalsegmental glomerulosclerosis (FSGS), endocrine ophthalmopathy,uveoretinitis, chorioretinitis, fibromyalgia, multiple endocrinefailure, Schmidt's syndrome, adrenalitis, gastric atrophy, preseniledementia, demyelinating diseases such as autoimmune demyelinatingdiseases, diabetic nephropathy, Dressler's syndrome, alopecia areata,CREST syndrome (calcinosis, Raynaud's phenomenon, esophagealdysmotility, sclerodactyly, and telangiectasia), male and femaleautoimmune infertility, mixed connective tissue disease, Chagas'disease, rheumatic fever, recurrent abortion, farmer's lung, erythemamultiforme, post-cardiotomy syndrome, Cushing's syndrome, bird-fancier'slung, allergic granulomatous angiitis, benign lymphocytic angiitis,Alport's syndrome, alveolitis such as allergic alveolitis and fibrosingalveolitis, interstitial lung disease, transfusion reaction, leprosy,malaria, leishmaniasis, kypanosomiasis, schistosomiasis, ascariasis,aspergillosis, Sampter's syndrome, Caplan's syndrome, dengue,endocarditis, endomyocardial fibrosis, diffuse interstitial pulmonaryfibrosis, interstitial lung fibrosis, idiopathic pulmonary fibrosis,fibrosis of any organ or tissue, cystic fibrosis, endophthalmitis,erythema elevatum et diutinum, erythroblastosis fetalis, eosinophilicfaciitis, Shulman's syndrome, Felty's syndrome, flariasis, cyclitis suchas chronic cyclitis, heterochronic cyclitis, iridocyclitis, or Fuch'scyclitis, Henoch-Schonlein purpura, human immunodeficiency virus (HIV)infection, echovirus infection, cardiomyopathy, Alzheimer's disease,parvovirus infection, rubella virus infection, post-vaccinationsyndromes, congenital rubella infection, Epstein-Barr virus infection,mumps, Evan's syndrome, autoimmune gonadal failure, Sydenham's chorea,post-streptococcal nephritis, thromboangitis ubiterans, thyrotoxicosis,tabes dorsalis, chorioiditis, giant cell polymyalgia, endocrineophthamopathy, chronic hypersensitivity pneumonitis,keratoconjunctivitis sicca, epidemic keratoconjunctivitis, idiopathicnephritic syndrome, minimal change nephropathy, benign familial andischemia-reperfusion injury, retinal autoimmunity, joint inflammation,bronchitis, chronic obstructive airway disease, silicosis, aphthae,aphthous stomatitis, arteriosclerotic disorders, aspermiogenese,autoimmune hemolysis, Boeck's disease, cryoglobulinemia, Dupuytren'scontracture, endophthalmia phacoanaphylactica, enteritis allergica,erythema nodosum leprosum, idiopathic facial paralysis, chronic fatiguesyndrome, febris rheumatica, Hamman-Rich's disease, sensoneural hearingloss, haemoglobinuria paroxysmatica, hypogonadism, ileitis regionalis,leucopenia, mononucleosis infectiosa, traverse myelitis, primaryidiopathic myxedema, nephrosis, ophthalmia symphatica, orchitisgranulomatosa, pancreatitis, polyradiculitis acuta, pyodermagangrenosum, Quervain's thyreoiditis, acquired spenic atrophy,infertility due to antispermatozoan antibodies, non-malignant thymoma,vitiligo, SCID and Epstein-Barr virus-associated diseases, acquiredimmune deficiency syndrome (AIDS), parasitic diseases such asLeishmania, toxic-shock syndrome, food poisoning, conditions involvinginfiltration of T cells, leukocyte-adhesion deficiency, immune responsesassociated with acute and delayed hypersensitivity mediated by cytokinesand T-lymphocytes, diseases involving leukocyte diapedesis, multipleorgan injury syndrome, antigen-antibody complex-mediated diseases,antiglomerular basement membrane disease, allergic neuritis, autoimmunepolyendocrinopathies, oophoritis, primary myxedema, autoimmune atrophicgastritis, sympathetic ophthalmia, rheumatic diseases, mixed connectivetissue disease, nephrotic syndrome, insulitis, polyendocrine failure,peripheral neuropathy, autoimmune polyglandular syndrome type I,adult-onset idiopathic hypoparathyroidism (AOIH), alopecia totalis,dilated cardiomyopathy, epidermolisis bullosa acquisita (EBA),hemochromatosis, myocarditis, nephrotic syndrome, primary sclerosingcholangitis, purulent or nonpurulent sinusitis, acute or chronicsinusitis, ethmoid, frontal, maxillary, or sphenoid sinusitis, aneosinophil-related disorder such as eosinophilia, pulmonary infiltrationeosinophilia, eosinophilia-myalgia syndrome, Loffler's syndrome, chroniceosinophilic pneumonia, tropical pulmonary eosinophilia,bronchopneumonic aspergillosis, aspergilloma, or granulomas containingeosinophils, anaphylaxis, seronegative spondyloarthritides,polyendocrine autoimmune disease, sclerosing cholangitis, sclera,episclera, chronic mucocutaneous candidiasis, Bruton's syndrome,transient hypogammaglobulinemia of infancy, Wiskott-Aldrich syndrome,ataxia telangiectasia, autoimmune disorders associated with collagendisease, rheumatism, neurological disease, ischemic re-perfusiondisorder, reduction in blood pressure response, vascular dysfunction,antgiectasis, tissue injury, cardiovascular ischemia, hyperalgesia,cerebral ischemia, and disease accompanying vascularization, allergichypersensitivity disorders, glomerulonephritides, reperfusion injury,reperfusion injury of myocardial or other tissues, dermatoses with acuteinflammatory components, acute purulent meningitis or other centralnervous system inflammatory disorders, ocular and orbital inflammatorydisorders, granulocyte transfusion-associated syndromes,cytokine-induced toxicity, acute serious inflammation, chronicintractable inflammation, pyelitis, pneumonocirrhosis, diabeticretinopathy, diabetic large-artery disorder, endarterial hyperplasia,peptic ulcer, valvulitis, and endometriosis. Other examples ofautoimmune diseases may be disclosed elsewhere herein.

The term “cancer” as used herein is defined as disease characterized bythe rapid and uncontrolled growth of aberrant cells. Cancer cells canspread locally or through the bloodstream and lymphatic system to otherparts of the body. Examples of various cancers include but are notlimited to, breast cancer, prostate cancer, ovarian cancer, cervicalcancer, skin cancer, pancreatic cancer, colorectal cancer, bladdercancer, renal cancer, brain cancer, lymphoma, leukemia, lung cancer andthe like.

As used herein, “combination” as in the phrase “a first agent incombination with a second agent” includes co-administration of a firstagent and a second agent, which for example may be dissolved orintermixed in the same pharmaceutically acceptable carrier, oradministration of a first agent, followed by the second agent, oradministration of the second agent, followed by the first agent.

The term “concomitant” as in the phrase “concomitant therapeutictreatment” includes administering an agent in the presence of a secondagent. A concomitant therapeutic treatment method includes methods inwhich the first, second, third, or additional agents areco-administered. A concomitant therapeutic treatment method alsoincludes methods in which the first or additional agents areadministered in the presence of a second or additional agents, whereinthe second or additional agents, for example, may have been previouslyadministered. A concomitant therapeutic treatment method may be executedstep-wise by different actors. For example, one actor may administer toa subject a first agent and a second actor may administer to the subjecta second agent, and the administering steps may be executed at the sametime, or nearly the same time, or at distant times, so long as the firstagent (and additional agents) are after administration in the presenceof the second agent (and additional agents). The actor and the subjectmay be the same entity (e.g., a human).

The term “combination therapy”, as used herein, refers to theadministration of two or more therapies, two or more therapeutic agents,or a combination of at least one therapeutic agent and at least onemethod of therapy, such as radiation, immunotherapy, and chemotherapy.

A “disease” is a state of health of an animal wherein the animal cannotmaintain homeostasis, and wherein if the disease is not ameliorated thenthe animal's health continues to deteriorate. In contrast, a “disorder”in an animal is a state of health in which the animal is able tomaintain homeostasis, but in which the animal's state of health is lessfavorable than it would be in the absence of the disorder. Leftuntreated, a disorder does not necessarily cause a further decrease inthe animal's state of health.

A disease or disorder is “alleviated” if the severity of a sign orsymptom of the disease or disorder, the frequency with which such a signor symptom is experienced by a patient, or both, is reduced.

The term “human microbiome” refers to the totality of microbes, theirgenetic elements (genomes), and environmental interactions in the humanbody.

The term “immune response” encompasses both cellular and humoral immuneresponses, including stimulating the production of cytokines,stimulating the proliferation of immune cells, stimulating theactivation of immune cells, or stimulating the lytic activity of immunecells. Examples of immune responses stimulated by the methods of theinvention are the secretion of cytokines, the activation of NK cells,the proliferation of B cells, T cells, macrophages, monocytes, and otherimmune cells, and other immune responses. To detect a cellular immuneresponse, for example, T cell effector activity against cells expressingthe antigen can be detected using standard assays, e.g., target-cellkilling, macrophage activation, B-cell activation or lymphokineproduction. Humoral responses can be measured by detecting theappearance of, or the increase in titer of, for example,antigen-specific antibodies, using methods known in the art, such asELISA. The progress of the antibody response can be determined bymeasuring class switching, such as the switch from an early IgM responseto a later IgG response.

The term “inhibit,” as used herein, means to suppress or block anactivity or function by at least about ten percent relative to a controlvalue. As an example, the activity is suppressed or blocked by 50%compared to a control value, by 75%, or by 95% or more.

The term “psoriasis” as used herein includes at least seven types ofpsoriasis: plaque psoriasis, guttate psoriasis, inverse psoriasis,pustular psoriasis, psoriasis vulgaris, seborrheic psoriasiserythrodermic psoriasis, nail psoriasis, and psoriatic arthritis.

The term “regulatory T cells” or “Tregs” refers to T cells that suppressan abnormal or excessive immune response and play a role in immunetolerance. The regulatory T cells are typically transcription factorFoxp3-positive CD4-positive T cells. The regulatory T cells of thepresent invention also include transcription factor Foxp3-negativeregulatory T cells that are IL-10-producing CD4-positive T cells.

The term “induces proliferation or accumulation of regulatory T cells”refers to an effect of inducing the differentiation of immature T cellsinto regulatory T cells, which differentiation leads to theproliferation and/or the accumulation of regulatory T cells. Further,the meaning of “induces proliferation or accumulation of regulatory Tcells” includes in-vivo effects, in vitro effects, and ex vivo effects.

The term “uveitis” as used herein refers to inflammation of the eye thatmay affect the uvea, or middle layer of the eye but also the lens,retina, optic nerve, and vitreous chamber. Uveitis (pronouncedyou-vee-EYE-tis) may be inflammation of the uvea, the middle layer ofthe eye that includes the iris, ciliary body and choroid. Uveitis can becaused by (1) autoimmune disorders (due to aberrant T cell-mediatedresponses triggered by inflammation and directed against retinal orcross-reactive antigens), (2) infections, (3) trauma, or (4) could beidiopathic (unknown cause). The type of uveitis is classified by whereinflammation occurs in the uvea. Anterior uveitis is inflammation of theiris (iritis) or the iris and ciliary body. Intermediate uveitis isinflammation of the ciliary body. Posterior uveitis is inflammation ofthe choroid. Diffuse uveitis (also called panuveitis) is inflammation ofall areas of the uvea. Uveitis may involve the full eye (panuveitis) ora segment of the eye (anterior, intermediate or posterior). Examples ofuveitis include, but are not limited to, anterior uveitis (comprisingiritis, iridiocyclitis, and anterior cylitis), intermediate uveitis(comprising pars planitis, posterior cyclitis, and hyalitis), posterioruveitis (comprising focal, multifocal or diffuse choroiditis,chorioretinitis, retinochoroiditis, retinitis, and neuroretinitis),panuveitis, acute uveitis, recurring uveitis and chronic uveitis. In oneembodiment, uveitis is non-infectious uveitis. Examples of causes ofnon-infectious uveitis include, but are not limited to systemicautoimmune disorders (such as, for example Behcet's disease andVogt-Koyanagi-Harada (VKH) disease); trauma and surgery. In one example,non-infectious uveitis is idiopathic non-infectious uveitis.

The terms “tolerance” and “immune tolerance” refer to the process bywhich the immune system does not attack an antigen.

The terms “tolerance induction” or “inducing tolerance” refer to aprocess by which tolerance to external antigens can be created bymanipulating the immune system.

The terms “treatment”, “treating” and the like are used herein togenerally mean obtaining a desired pharmacological and/or physiologicaleffect. The effect may be prophylactic in terms of completely orpartially preventing a disease or disorder, or a symptom thereof, and/ormay be therapeutic in terms of partially or completely curing a diseaseor disorder, and/or adverse effect attributed to the disease ordisorder. The term “treatment” as used herein includes any treatment ofa disease or disorder in a subject and includes: (a) preventing adisease or disorder from occurring in a subject which may be predisposedto the disease or disorder; (b) inhibiting the disease or disorder, i.e.arresting its development: or (c) relieving the disease or disorder,i.e. causing regression of the disease or disorder.

The terms “effective amount” and “pharmaceutically effective amount”refer to a sufficient amount of an agent to provide a desired biologicalresult. That result can be reduction and/or alleviation of a sign,symptom, or cause of a disease or disorder, or any other desiredalteration of a biological system.

The term “leukemia,” as used herein, may refer to at least one of thefollowing types of leukemia: myeloproliferative neoplasms (MPNs),polycythemia vera (PV), essential thrombocytosis (ET),idiopathic/myelofibrosis (MF), acute myeloid leukemia (AML), andchildhood acute lymphoblastic leukemia (ALL). The term “leukemia” mayalso refer generally to cancer of blood forming tissue, or to bloodcancer.

The term “microbiota” refers, collectively, to the entirety of microbesfound in association with a higher organism, such as a human.

A “therapeutically effective amount” refers to that amount whichprovides a therapeutic effect for a given condition and administrationregimen. In particular, “therapeutically effective amount” means anamount that is effective to prevent, alleviate or ameliorate symptoms ofa disease or disorder, or prolong the survival of the subject beingtreated, which may be a human or non-human animal.

The term “macular degeneration” as used herein refers to any and allforms of macular degeneration, including “wet” and “dry” maculardegeneration. There are two types of age-related macular degeneration(AMD): the dry (atrophic) form and the wet (exudative) form. The dryform of AMD affects about 90 percent of AMD patients and usually beginswith the formation of tiny yellow deposits called drusen in the macula.Drusen usually do not cause serious loss of vision, but can causedistortion of vision. However, for reasons that are not yet understood,sometimes drusen will cause the macula to thin and break down, slowlyleading to vision loss. The wet form of AMD occurs in about 10 percentof AMD patients. It is caused by the growth of abnormal blood vesselsbeneath the macula that can leak fluid and blood to form exudate. Thewet form of AMD typically causes significant vision problems in theaffected eye and can progress very rapidly, causing permanent centralvision loss.

As used herein, the term “pharmaceutical composition” refers to amixture of at least one compound of the invention with other chemicalcomponents and entities, such as carriers, stabilizers, diluents,dispersing agents, suspending agents, thickening agents, and/orexcipients. The pharmaceutical composition facilitates administration ofthe compound to an organism. Multiple techniques of administering acompound exist in the art including, but not limited to, intravenous,oral, aerosol, parenteral, ophthalmic, pulmonary and topicaladministration.

“Pharmaceutically acceptable” refers to those properties and/orsubstances which are acceptable to the patient from apharmacological/toxicological point of view and to the manufacturingpharmaceutical chemist from a physical/chemical point of view regardingcomposition, formulation, stability, patient acceptance andbioavailability.

As used herein, the term “pharmaceutically acceptable carrier” means apharmaceutically acceptable material, composition or carrier, such as aliquid or solid filler, stabilizer, dispersing agent, suspending agent,diluent, excipient, thickening agent, solvent, or encapsulatingmaterial, involved in carrying or transporting a compound useful withinthe invention within or to the patient such that it may perform itsintended function. Typically, such constructs are carried or transportedfrom one organ, or portion of the body, to another organ, or portion ofthe body. Each carrier must be “acceptable” in the sense of beingcompatible with the other ingredients of the formulation, including thecompound useful within the invention, and not injurious to the patient.Some examples of materials that may serve as pharmaceutically acceptablecarriers include: sugars, such as lactose, glucose and sucrose;starches, such as corn starch and potato starch; cellulose, and itsderivatives, such as sodium carboxymethyl cellulose, ethyl cellulose andcellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients,such as cocoa butter and suppository waxes; oils, such as peanut oil,cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; glycols, such as propylene glycol; polyols, such asglycerin, sorbitol, mannitol, polyethylene glycol and glycerol; esters,such as ethyl oleate and ethyl laurate; agar; buffering agents, such asmagnesium hydroxide and aluminum hydroxide; surface active agents;alginic acid; pyrogen-free water; isotonic saline; Ringer's solution;ethyl alcohol; phosphate buffer solutions; and other non-toxiccompatible substances employed in pharmaceutical formulations. As usedherein, “pharmaceutically acceptable carrier” also includes any and allcoatings, antibacterial and antifungal agents, and absorption delayingagents, and the like that are compatible with the activity of thecompound useful within the invention, and are physiologically acceptableto the patient. Supplementary active compounds may also be incorporatedinto the compositions. The “pharmaceutically acceptable carrier” mayfurther include a pharmaceutically acceptable salt of the compounduseful within the invention. Other additional ingredients that may beincluded in the pharmaceutical compositions used in the practice of theinvention are known in the art and described, for example in Remington'sPharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton,Pa.), which is incorporated herein by reference.

The term “neonate” as used herein generally refers to an infant or ayoung mammal, e.g., a human being.

The term “nutritional composition” may refer to a food product intendedfor human consumption, for example, a beverage, a drink, a bar, a snack,an ice cream, a dairy product, for example a chilled or a shelf-stabledairy product, a fermented dairy product, a drink, for example amilk-based drink, an infant formula, a growing-up milk, a confectioneryproduct, a chocolate, a cereal product such as a breakfast cereal, asauce, a soup, an instant drink, a frozen product intended forconsumption after heating in a microwave oven or a conventional oven, aready-to-eat product, a fast food or a nutritional formula.

The terms “patient,” “subject,” “individual,” and the like are usedinterchangeably herein, and refer to any animal, or cells thereofwhether in vitro or in situ, amenable to the methods described herein.The patient, subject, or individual may be a human or a non-humananimal.

As used herein, the term “container” includes any receptacle for holdingthe pharmaceutical composition. For example, in one embodiment, thecontainer is the packaging that contains the pharmaceutical composition.In other embodiments, the container is not the packaging that containsthe pharmaceutical composition, i.e., the container is a receptacle,such as a box or vial that contains the packaged pharmaceuticalcomposition or unpackaged pharmaceutical composition and theinstructions for use of the pharmaceutical composition. Moreover,packaging techniques are well known in the art. It should be understoodthat the instructions for use of the pharmaceutical composition may beincluded on the packaging containing the pharmaceutical composition, andas such the instructions form an increased functional relationship tothe packaged product. However, it should be understood that theinstructions may contain information pertaining to the compound'sability to perform its intended function, e.g., treating or preventing adisease in a subject.

“Instructional material,” as that term is used herein, includes apublication, a recording, a diagram, or any other medium of expressionwhich can be used to communicate the usefulness of components of theinvention in a kit for identifying or alleviating or treating thevarious diseases or disorders recited herein. Optionally, oralternately, the instructional material may describe one or more methodsof identifying or alleviating the diseases or disorders in a cell or atissue of a subject. The instructional material of the kit may, forexample, be affixed to a container that contains the compositions of theinvention or be shipped together with a container that contains thecompositions of the invention. Alternatively, the instructional materialmay be shipped separately from the container with the intention that therecipient uses the instructional material and the compositioncooperatively.

The term “lymphoma,” as that term is used herein, may refer to at leastone of the following types of lymphoma: activated B-cell-like diffuselarge B cell lymphoma (ABC DLBCL), follicular lymphoma (FL), mucosaassociated lymphoid tissue lymphoma (MALT), Hodgkin lymphoma (HL), andprimary mediastinal B cell lymphoma (PMBL).

“Short chain fatty acids” (SCFA) are fatty acids typically withaliphatic tails shorter than aliphatic tails of long chain fatty acids.As used herein, the term short chain fatty acid may also refer to saltsor esters of fatty acids, especially pharmaceutically acceptable saltsand esters of fatty acids (e.g., sodium butyrate, arginine butyrate).

As used herein, the term “stimulate an immune response” includesstimulating, eliciting, increasing, enhancing, sustaining, and/orimproving the stimulation of new immune response or of a preexistingimmune response. Thus, “stimulating an immune response” as animmunotherapy refers to enhancing the therapeutic efficacy, increasingsurvival time, slowing the progression of a cancerous tumor or shrinkingthe cancerous tumor size, preventing the spread of a tumor or ofmetastases, preventing or slowing the recurrence of treated cancer,eliminating cancer cells not killed by earlier treatments, targetingpotential cancer cells or targeting antigens derived from a virusassociated with cancer. In the methods of this invention, theimmunotherapeutic agent and the compound of selected from formulae (I),(II), (III), (IV) and (V) are administered in an amount effective tostimulate an immune response in the subject individual at a dosesufficient to generate an effective immune response without unacceptabletoxicity. As will be understood by one of skill in the art, themagnitude of the immune response and the maintenance of that responsemay have varying degrees which will be recognized a having a potentialtherapeutic or prophylactic benefit.

The term “vasculitis,” as that term is used herein, may refer to atleast one of the following types of vasculitis: giant cell arteritis,Takayasu disease, Churg-Strauss syndrome, Wegener granulomatosis,microscopic polyangiitis, essential cryoglobulinemic vasculitis,Henoch-Shoenlein purpura, Kawasaki disease, polyarteritis nodosa,anti-neutrophil cytoplasmic antibody vasculitis, and necrotizing andcresentic glomerulonephritis.

Ranges: throughout this disclosure, various aspects of the invention canbe presented in a range format. It should be understood that thedescription in range format is merely for convenience and brevity andshould not be construed as an inflexible limitation on the scope of theinvention. Accordingly, the description of a range should be consideredto have specifically disclosed all the possible subranges as well asindividual numerical values within that range. For example, descriptionof a range such as from 1 to 6 should be considered to have specificallydisclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numberswithin that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. Thisapplies regardless of the breadth of the range.

Description

The present invention is based, in part upon the discovery that shortchain fatty acids (SCFA) serve as modulators of multiple cellularsignaling proteins, including, but not limited to, IL-18, TLR3, IFN-γ,TNFα, TGF-β, MyD88, PI3K/Akt, JAK/STAT, Smad 2/3, Smad 4, IL-10, Notch,hedgehog, Wnt (beta-catenin), matrix metallo-proteinases 9 and 10,tissue inhibitor of metalloproteinases, nodal and NF-κB signaling. Invarious embodiments, the signaling proteins that are modulated by SCFAsthemselves modulate biological pathways or processes including, but notlimited to, inflammation, immunity, proliferation, differentiation,apoptosis, oncogenesis, transcription of DNA, cytokine production, cellsurvival, angiogenesis, fibrogenesis and cellular responses to stimulisuch as stress, cytokines, free radicals, heavy metals, and ultravioletirradiation

In one embodiment, the invention relates to compositions comprising atleast one SCFA and methods of use for treating or preventing medicaldiseases or disorders characterized by elevated levels or abnormalexpression of at least one of IL-18, TLR3, IFN-γ, TNFα, TGF-β, MyD88,PI3K/Akt, JAK/STAT, Smad 2/3, Smad 4 or IL-10 signaling. In oneembodiment, the invention relates to compositions comprising at leastone SCFA and methods of use for treating or preventing medical diseasesor disorders characterized by decreased levels or abnormal expression ofNF-κB signaling.

Compositions

In one embodiment, the invention provides compositions comprising atleast one short chain fatty acid (SCFA), SCFA precursor, SCFAbiosynthesis precursor, a derivative thereof, a SCFA moiety or acombination thereof.

In one embodiment, the composition of the invention comprises at leastone SCFA, or a compound comprising a SCFA moiety. In one embodiment, aSCFA or SCFA moiety includes, but is not limited to, acetic acid,butyric acid (BA), C3-C12 fatty acids, C3-C10 fatty acids, C3-C8 fattyacids, methoxyacetic acid, valproic acid (VPA), propionic acid,3-methoxypropionic acid, ethoxyacetic acid, formic acid, isobutyricacid, tributyrin, N-acetylbutyrate (as well as other forms of butyrate,e.g., phenylbutyrate, isobutyrate, pivaloyloxymethyl butyrate,monoacetone glucose 3-butyrate), isovaleric acid, valeric acid,isocaproic acid, caproic acid, lactic acid, succinic acid, pyruvic acid,octanoic acid, dodecanoic acid, (4R)-4-hydroxypentanoic acid,2-ethylhydracrylic acid, 2-hydroxy-3-methylpentanoate,2-hydroxy-3-methylpentanoic acid, 2-methylbut-2-enoic acid,2-oxobutanoic acid, 3-hydroxypentanoic acid, 3-methylbut-2-enoic acid,butenoic acid, methylbutyric acid, dimethylbutyric acid, pentadienoicacid, pentenoic acid, pivalic acid, propynoic acid and a combinationthereof. In one embodiment, the SCFA or SCFA moiety includes compoundsor structures with at least 12 carbon atoms, at least 11 carbon atoms,at least 10 carbon atoms, at least 9 carbon atoms, at least 8 carbonatoms, at least 7 carbon atoms, at least 6 carbon atoms, at least 5carbon atoms, at least 4 carbon atoms, at least 3 carbon atoms, and atleast 2 carbon atoms. In one embodiment, the SCFA or SCFA moietyincludes compounds or structures with less than 13 carbon atoms, lessthan 12 carbon atoms, less than 11 carbon atoms, less than 10 carbonatoms, less than 9 carbon atoms, less than 8 carbon atoms, or less than7 carbon atoms. In one embodiment, the SCFA or SCFA moiety is not abranched fatty acid. In one embodiment, the SCFA or SCFA moiety is abranched fatty acid.

In one embodiment, the composition of the invention comprises at leastone compound comprising a precursor of a SCFA, or a moiety thereof. Inone embodiment, the precursor, or moiety thereof, is selected from thegroup including, but not limited to, plant cell-wall polysaccharides,dietary nonstarch polysaccharides (NSP) a salt of lactate, a salt ofsuccinate, a salt of formate, 1,2-propenedol, trypamine, indole,indole-3-acetate, and a combination thereof.

In one embodiment, the composition of the invention comprises at leastone compound comprising a biosynthesis precursor of a SCFA, or a moietythereof. In one embodiment, the biosynthesis precursor, or moietythereof, is selected from the group including, but not limited to anacetyl-CoA carboxylase inhibitor, an adenosine monophosphate kinase(AMPK) activator, vitamin D, and a combination thereof.

In one embodiment, the composition of the invention comprises a salt ofa SCFA, or a derivative thereof. For example, a salt of butyric acid maybe one or more of sodium butyrate, magnesium butyrate or calciumbutyrate. In one embodiment, the composition comprises one or more ofmagnesium butyrate and calcium butyrate.

In one embodiment, the composition of the invention comprises aderivative of a SCFA. In one embodiment, the derivative comprises atleast one SCFA moiety linked to at least one additional moiety. In oneembodiment, the derivative comprises at least one SCFA moiety linked toat least one polyethylene glycol (PEG) moiety. In one embodiment, the atleast one SCFA moiety linked to at least one PEG moiety hydrolyzes undera low pH condition to yield at least one SCFA molecule and at least onePEG molecule.

In one embodiment, the composition of the invention comprises acombination of SCFAs, and/or derivatives thereof. In one embodiment, thecomposition is prepared at amounts of at least 10 mM, at least 20 mM, atleast 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, or moreof each or all the compounds of the composition.

Derivatives of SCFAs, e.g., having substituents on the carbon chain suchas O, S, N, methyl, ethyl, halogen, and other groups that do notinterfere with the compound's therapeutic activity may also be used toform the compositions of this invention. In one embodiment, the compoundof the invention comprises at least one SCFA linked to at least oneadditional moiety, such as O, S, N, methyl, ethyl, halogen, and othergroups that do not interfere with the compound's therapeutic activity.

In some instances, the SCFA of the invention can be pegylated.Polyethylene glycol (PEG) has been widely used in biomaterials,biotechnology and medicine primarily because PEG is a biocompatible,nontoxic, nonimmunogenic and water-soluble polymer (Zhao and Harris, ACSSymposium Series 680: 458-72, 1997). In the area of drug delivery, PEGderivatives have been widely used in covalent attachment (i.e.,“PEGylation”) to proteins to reduce immunogenicity, proteolysis andkidney clearance and to enhance solubility (Zalipsky, Adv. Drug Del.Rev. 16:157-82, 1995). Similarly, PEG has been attached to low molecularweight, relatively hydrophobic drugs to enhance solubility, reducetoxicity and alter biodistribution. Typically, PEGylated drugs areinjected as solutions. However, they may be administered orally, or byanother route.

A closely related application is synthesis of crosslinked degradable PEGnetworks or formulations for use in drug delivery since much of the samechemistry used in design of degradable, soluble drug carriers can alsobe used in design of degradable gels (Sawhney et al., Macromolecules 26:581-87, 1993). It is also known that intermacromolecular complexes canbe formed by mixing solutions of two complementary polymers. Suchcomplexes are generally stabilized by electrostatic interactions(polyanion-polycation) and/or hydrogen bonds (polyacid-polybase) betweenthe polymers involved, and/or by hydrophobic interactions between thepolymers in an aqueous surrounding (Krupers et al., Eur. Polym J.32:785-790, 1996). For example, mixing solutions of polyacrylic acid(PAAc) and polyethylene oxide (PEO) under the proper conditions resultsin the formation of complexes based mostly on hydrogen bonding.Dissociation of these complexes at physiologic conditions has been usedfor delivery of free drugs (i.e., non-PEGylated). In addition, complexesof complementary polymers have been formed from both homopolymers andcopolymers.

In one embodiment, the composition of the invention comprises aprecursor of a SCFA alone or in combination with one or more SCFAs.Precursors of SCFAs include, but are not limited to, a salt of formate,a salt of lactate, a salt of succinate, 1,2-propenedol, trypamine,indole, and indole-3-acetate.

In one embodiment, the composition of the invention comprises aprecursor of SCFA biosynthesis alone or in combination with one or moreSCFA. Precursors of SCFA biosynthesis include, but are not limited to, asalt of formate, a salt of lactate, a salt of succinate, acetyl-CoAcarboxylase inhibitors, adenosine monophosphate kinase (AMPK)activators, and vitamin D.

SCFAs stimulate T regulatory (Treg) cell function, which accounts forsome of their anti-inflammatory properties. Given that inhibitors ofacetyl-CoA carboxylase also promote Treg cell function, in oneembodiment, a composition comprises an inhibitor of acetyl-CoAcarboxylase, including but not limited to, biotin or its naturally orchemically synthesized analogs (which are acetyl-CoA carboxylaseinhibitors) alone or in combination with one or more other SCFAs, tostimulate Treg functions.

In various embodiments, a compound comprising at least one SCFA, or acompound comprising a SCFA moiety of the invention may be combined withone or more compounds, such as one or more additional therapeutic agent,for a particular disease or disorder. In one embodiment, the one or moreSCFAs of the invention may be in the same composition as one or moreadditional therapeutic agent. In various embodiments the composition maycomprise at least 1, at least 2, at least 3, at least 4, at least 5, atleast 6, at least 7, at least 8, at least 9, at least 10 or more than 10additional therapeutic agents. Exemplary additional therapeutic agentsand/or compounds that can be included in a composition of the inventionare discussed in detail elsewhere herein.

Methods

The present invention provides methods for treating or preventing amammalian disease in a subject in need thereof by administration to saidsubject a therapeutically effective amount of the compositions of thepresent invention.

General examples of target diseases for which the composition(s) is/areuseful for treatment (reducing adverse effects or prevention) includeautoimmune diseases, allergic diseases, infectious diseases, andrejection in organ transplantations, such as inflammatory bowel disease(IBD), ulcerative colitis, Crohn's disease, sprue, autoimmune arthritis,rheumatoid arthritis, Type I diabetes, multiple sclerosis, graft vs.host disease following bone marrow transplantation, osteoarthritis,juvenile chronic arthritis, Lyme arthritis, psoriatic arthritis,reactive arthritis, spondy loarthropathy, systemic lupus erythematosus,insulin dependent diabetes mellitus, thyroiditis, asthma, psoriasis,dermatitis scleroderma, atopic dermatitis, graft versus host disease,acute or chronic immune disease associated with organ transplantation,sarcoidosis, atherosclerosis, disseminated intravascular coagulation,Kawasaki's disease, Grave's disease, nephrotic syndrome, chronic fatiguesyndrome, Wegener's granulomatosis, Henoch-Schoenlejn purpurea,microscopic vasculitis of the kidneys, uveitis, septic shock, toxicshock syndrome, sepsis syndrome, cachexia, acquired immunodeficiencysyndrome, acute transverse myelitis, Huntington's chorea, Parkinson'sdisease, Alzheimer's disease, stroke, primary biliary cirrhosis,hemolytic anemia, polyglandular deficiency type I syndrome andpolyglandular deficiency type II syndrome, Schmidt's syndrome, adult(acute) respiratory distress syndrome, alopecia, alopecia areata,seronegative arthopathy, arthropathy, Reiter's disease, psoriaticarthropathy, chlamydia, yersinia and salmonella associated arthropathy,spondyloarhopathy, atheromatous disease/arteriosclerosis, allergiccolitis, atopic allergy, food allergies such as peanut allergy, tree nutallergy, egg allergy, milk allergy, soy allergy, wheat allergy, seafoodallergy, shellfish allergy, or sesame seed allergy, autoimmune bullousdisease, pemphigus vulgaris, pemphigus foliaceus, pemphigoid, linear IgAdisease, autoimmune haemolytic anaemia, Coombs positive haemolyticanaemia, acquired pernicious anaemia, juvenile pernicious anaemia,myalgic encephalitis/Royal Free Disease, chronic mucocutaneouscandidiasis, giant cell arteritis, Acquired Immunodeficiency DiseaseSyndrome, Acquired Immunodeficiency Related Diseases, common variedimmunodeficiency (common variable hypogammaglobulinaemia), dilatedcardiomyopathy, fibrotic lung disease, cryptogenic fibrosing alveolitis,postinflammatory interstitial lung disease, interstitial pneumonitis,connective tissue disease associated interstitial lung disease, mixedconnective tissue disease associated lung disease, systemic sclerosisassociated interstitial lung disease, rheumatoid arthritis associatedinterstitial lung disease, systemic lupus erythematosus associated lungdisease, dermatomyositis/polymyositis associated lung disease, Sjogren'sdisease associated lung disease, ankylosing spondylitis associated lungdisease, vasculitic diffuse lung disease, haemosiderosis associated lungdisease, drug-induced interstitial lung disease, radiation fibrosis,bronchiolitis obliterans, chronic eosinophilic pneumonia, lymphocyticinfiltrative lung disease, postinfectious interstitial lung disease,gouty arthritis, autoimmune mediated hypoglycemia, type B insulinresistance with acanthosis nigricans, hypoparathyroidism, acute immunedisease associated with organ transplantation, chronic immune diseaseassociated with organ transplantation, osteoarthrosis, primarysclerosing cholangitis, idiopathic leucopenia, autoimmune neutropenia,renal disease NOS, glomerulonephritides, microscopic vasulitis of thekidneys, discoid lupus, erythematosus, male infertility idiopathic orNOS, sperm autoimmunity, multiple sclerosis (all subtypes), insulindependent diabetes mellitus, sympathetic ophthalmia, pulmonaryhypertension secondary to connective tissue disease, Goodpasture'ssyndrome, pulmonary manifestation of polyarteritis nodosa, acuterheumatoid fever, rheumatoid spondylitis, Still's disease, systemicsclerosis, Takayasu's disease/arteritis, autoimmune thrombocytopenia,idiopathic thrombocytopenia, autoimmune thyroid disease,hyperthyroidism, goitrous autoimmune hypothyroidism (Hashimoto'sdisease), atrophic autoimmune hypothyroidism, primary myxoedema,phacogenic uveitis, primary vasculitis, vitiligo, allergic rhinitis(pollen allergies), anaphylaxis, pet allergies, latex allergies, drugallergies, allergic rhinoconjuctivitis, eosinophilic esophagitis,hypereosinophilic syndrome, eosinophilic gastroenteritis cutaneous lupuserythematosus, eosinophilic esophagitis, hypereosinophilic syndrome, andeosinophilic gastroenteritis, and diarrhea.

In one embodiment, the invention provides methods for the treatment orprevention of at least one disease or disorder in a subject, comprisingadministering to the subject at least one composition comprising a SCFAor a compound comprising a SCFA moiety, optionally in combination withat least one additional agent or therapy.

In some embodiments, the administered compositions of the presentinvention can increase the number of disease-free days, reduce theseverity of a disease or disorder, reduce the risk of developing adisease or disorder, reduce the risk of recurrence of a disease ordisorder, or a combination thereof in the subject. The administeredcompositions of the present invention can increase the number ofdisease-free days by 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%,40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%,54%, 55%, 56%, 57%, 58%, 59%, 60% or more in the subject as compared toa subject who is not receiving treatment. The administered compositionsof the present invention can reduce the severity of a disease ordisorder by 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%,42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60% or more in the subject as compared to a subjectwho is not receiving treatment. The administered compositions of thepresent invention can reduce the risk of developing a disease ordisorder by 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%,42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60% or more in the subject as compared to a subjectwho is not receiving treatment. The administered compositions of thepresent invention can reduce the risk of recurrence of a disease ordisorder by 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%,42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60% or more in the subject as compared to a subjectwho is not receiving treatment.

In some embodiments, diseases and disorders that can be treated,prevented or ameliorated include, but are not limited to, inflammatorydiseases and various cancer diseases. In some embodiments, theinflammatory diseases and disorders that can be treated or amelioratedinclude, but are not limited to, asthma, arthritis, allergic rhinitis,psoriasis, atopic dermatitis, inflammatory bowel diseases, Crohn'sdisease, an allergic or autoimmune disease or disorder associated withC-section delivery of a neonate, uveitis, and vasculitis. In someembodiments, the cancer diseases and disorders that can be treated orameliorated include, but are not limited to, leukemias and lymphomas.

Methods for treating exemplary diseases and disorders are providedbelow.

Skin Disorders

The present invention is partly based upon the discovery that SCFAs areeffective in the treatment of a skin disorder. In one embodiment, acombination of at least one SCFA with at least one other skin disordertreatment can be effective as a therapeutic approach for the treatmentof a skin disorder.

In one embodiment, the present invention provides methods for treatment,inhibition, prevention, or reduction of a skin disease or disorder byadministering a composition comprising a SCFA, as disclosed herein, to asubject in need thereof, optionally in combination with at least oneadditional agent or therapy. Skin diseases and disorders that may betreated using the methods of the invention include, but are not limitedto: psoriasis, plaque psoriasis, guttate psoriasis, inverse psoriasis,pustular psoriasis, psoriasis vulgaris, seborrheic psoriasis,erythrodermic psoriasis, nail psoriasis, psoriatic arthritis, systemiclupus erythematosus (SLE) rash, scleroderma (systemic sclerosis),diabetes related skin conditions, rheumatoid arthritis and associatedskin rashes (rheumatoid vasculitis), melanoma, vitiligo, eczema (atopicdermatitis), dyshidrotic eczema, rosacea, hives, impetigo, cellulitis,contact dermatitis, canker sores, acne, Lichen planus, actinickeratosis, ichthyosis vulgaris, dermatomyositis, and pemphigoid. In oneembodiment, the present invention provides methods for treatment,inhibition, prevention, or reduction of a skin disease or disorder,comprising administering a composition comprising a SCFA, as disclosedherein, to a subject in need thereof.

In one embodiment, a subject suffering from a skin disorder is a human.In one embodiment, a subject suffering from a skin disorder is anon-human animal.

In one embodiment, the invention relates to compositions comprising atleast one SCFA and at least one second compound for use as a therapeuticfor the treatment of skin disorders. In one embodiment, the SCFAcomprises one or more of formic acid, acetic acid, propionic acid,isobutyric acid, butyric acid, tributyrin, N-acetylbutyrate (as well asother forms of butyrate), isovaleric acid, valeric acid, isocaproicacid, caproic acid, lactic acid, succinic acid, pyruvic acid, octanoicacid, and dodecanoic acid. In one embodiment, the second compoundcomprises one or more of a PDE4 inhibitor, an anti-inflammatorycompound, a disease-modifying antirheumatic drug (DMARD), animmunosuppressant, a biologic agent, and a Cox-2 inhibitor.

In one embodiment, a composition for use in methods of treating skindisorders comprises 900 mg butyrate, 100 mg propionate, 10 mgapremilast, 10 mg magnesium, and 50 IU vitamin D3.

In one embodiment, an exemplary method for the treatment of vasculitiscomprises administration of a daily oral dose of at least 100 mg, atleast 200 mg, at least 300 mg, at least 400 mg, at least 500 mg, atleast 600 mg, at least 700 mg, at least 800 mg, at least 900 mg, atleast 1 g, at least 2 g, at least 3 g, at least 4 g, at least 5 g, atleast 6 g or more than 6 g of at least one SCFA, at least 1 time daily,at least 2 times daily, at least 3 times daily or more than 3 timesdaily, for at least one week, at least two weeks, at least three weeks,at least one month, at least 2 months, at least 3 months, at least 4months, at least 5 months, at least 6 months or for more than 6 months.

In one embodiment, an exemplary daily oral dosage for use in methods oftreating skin disorders comprises 3600 mg of butyrate, 400 mgpropionate, 40 mg Magnesium, and 200 IU Vitamin D3. In one embodiment,an exemplary dosage for use in methods of treating skin disorderscomprises 900-1800 mg of butyrate, 100-200 mg propionate, 10-20 mgMagnesium, and 50-100 IU Vitamin D3 administered 1-4 times daily. In oneembodiment, the composition is administered 1-4 times daily for at least1 week, at least 2 weeks, at least 3 weeks or for more than 3 week.

In one embodiment, an exemplary dosage for use in methods of treatingskin disorders comprises 1-2 g of butyrate, 100 mg propionate, 10-15 mgOtezla, 10-20 mg Magnesium, and 80-100 IU Vitamin D3.

In one embodiment, the composition comprising at least one SCFA is anenteric coated, extended release and sustained release capsule.

In one embodiment, a method of treating skin disorders comprisesadministration of an oral formulation of a SCFA in combination with atopical ointment. An exemplary topical ointment for use in treating skindisorder may comprise 40% clobetazol (0.05%) cream, 20% calcipotriene(vit D, 0.005%) cream, 20% vit E (0.5%) cream and 20% salicylic acid(10%) cream. An alternative exemplary topical ointment for use intreating skin disorder may comprise 40% clobetazol (0.05%) cream, 20%calcipotriene (vit D, 0.005%) cream, 20% vit E (0.5%) cream and 20% zinccream. In one embodiment zinc could be used together with salicylic acidin a topical ointment.

Eye Disease

In one embodiment, the invention relates to compositions comprising SCFAfor use as therapeutics for the treatment of eye diseases or disorders.In one embodiment, the eye disease or disorder is inflammatory. In oneembodiment, the inflammatory eye disease or disorder is uveitis. In oneembodiment, a composition comprises one or more SCFAs. In oneembodiment, the SCFAs include, but are not limited to, at least one offormic acid, acetic acid, propionic acid, isobutyric acid, butyric acid,tributyrin, N-acetylbutyrate (as well as other forms of butyrate),isovaleric acid, valeric acid, isocaproic acid, caproic acid, lacticacid, succinic acid, pyruvic acid, octanoic acid, and dodecanoic acid.In one embodiment, a composition comprises one or more of magnesium andcalcium salts of one or more of the compounds disclosed herein.

In one embodiment, the invention comprises a formulated compositioncomprising a SCFA or a compound comprising a SCFA moiety. In oneembodiment, the composition comprises an oral pharmaceutical or dietarycomposition comprising a SCFA or a compound comprising a SCFA moiety.

In one embodiment, the invention relates to a method of treating an eyedisease or disorder comprising administering to a subject a compositioncomprising a SCFA. In one embodiment, the eye disease or disorder isinflammatory. In one embodiment, the inflammatory eye disorder isuveitis. In one embodiment, the composition is administered orally. Inone embodiment, the composition is administered topically (e.g., in acream). In one embodiment, a subject suffering from an eye disorder is ahuman. In one embodiment, a subject suffering from an eye disorder is anon-human animal.

In one embodiment, the present invention provides methods for treatment,inhibition, prevention, or reduction of an eye disease or disorder byadministering a composition comprising a SCFA, as disclosed herein, to asubject in need thereof, optionally in combination with at least oneadditional agent or therapy. Eye diseases and disorders that may betreated using the methods of the invention include, but are not limitedto uveitis, macular degeneration, age related macular degeneration(AMD), inflammation following a surgical procedure (e.g., a cataractsurgery), Behçet's Disease of the eye, Sjogren's syndrome associateddiseases or disorders (e.g., dry eyes), blepharitis associated diseasesor disorders (e.g., rosacea) and any eye diseases or disordersassociated with other diseases discussed in detail elsewhere herein. Inone embodiment, the present invention provides methods for treatment,inhibition, prevention, or reduction of a disease associated with an eyedisease or disorder, comprising administering a composition comprising aSCFA or a compound comprising a SCFA moiety, as disclosed herein, to asubject in need thereof. The present invention is partly based on thediscovery that SCFA can be effective as a therapeutic approach for thetreatment of an eye disease or disorder, including inflammatory eyediseases or disorders.

In one embodiment, an exemplary method for the treatment or preventionof eye diseases or disorders comprises administration of a daily oraldose of a composition comprising at least 100 mg, at least 200 mg, atleast 300 mg, at least 400 mg, at least 500 mg, at least 600 mg, atleast 700 mg, at least 800 mg, at least 900 mg, at least 1 g, at least 2g, at least 3 g, least 4 g, at least 5 g, at least 6 g or more than 6 gof at least one SCFA at least 1 time daily, at least 2 times daily, atleast 3 times daily or more than 3 times daily, for at least one week,at least two weeks, at least three weeks, at least one month, at least 2months, at least 3 months, at least 4 months, at least 5 months, atleast 6 months or for more than 6 months.

In one embodiment, the composition comprising at least one SCFA is anenteric coated, extended release capsule. In one embodiment, anexemplary daily oral dose is 4-5 g of butyrate and 1.5-2 g of propionatein enteric coated, extended release capsules, twice/day.

In one embodiment, an exemplary method for the treatment or preventionof eye diseases or disorder comprises administration of a daily topicaldose of a composition comprising at least 100 nM, at least 200 nM, atleast 300 nM, at least 400 nM, at least 500 nM, at least 600 nM, atleast 700 nM, at least 800 nM, at least 900 nM, at least 1 μM, at least2 μM, at least 3 μM, least 4 μM, at least 5 μM, at least 6 μM, at least7 μM, at least 8 μM, at least 9 μM, at least 10 μM, at least 15 μM, atleast 20 μM, at least 25 μM, at least 30 μM, at least 35 μM, at least 40μM, at least 45 μM, at least 50 μM, or more than 50 μM of at least oneSCFA at least 1 time daily, at least 2 times daily, at least 3 timesdaily or more than 3 times daily, for at least one week, at least twoweeks, at least three weeks, at least one month, at least 2 months, atleast 3 months, at least 4 months, at least 5 months, at least 6 monthsor for more than 6 months.

In one embodiment, the composition comprising at least one SCFA is aneyedrop formulation. In one embodiment, an exemplary eyedrop formulationcontaining 20-30 μM of butyrate and 10-20 μM propionate can beadministered at least twice/day in combination with lubricants.

In one embodiment, eyedrops should be given four times/day (in case ofdisease flares), and twice/day up to few months, in combination withoral doses of SCFAs.

In one embodiment, the method for the treatment or prevention of eyediseases or disorder includes administering a combination of SCFAs andantibiotics and/or steroids. For example, in case of infectious uveitis:during first 3-5 days, use eyedrops (or eye injection) withantibiotics/steroids (at the doses prescribed by doctors), then duringfollowing 2-3 weeks, use eyedrops containing a mixture of SCFAs (20-30μM) and antibiotics/steroids (at half of the prescribed doses), thencontinue eyedrops with SCFAs (20-30 μM) only.

In one embodiment, the method for the treatment or prevention of eyediseases or disorder includes administering a combination ofcompositions comprising at least one SCFA. For example, in oneembodiment, the method includes a combination of at least one SCFAformulated for oral administration and at least one SCFA formulated foruse as eyedrops.

Allergy, Autoimmune and Asthma

The present invention contemplates the treatment or prevention ofallergies, autoimmune disease and asthma. The present invention ispartly based upon the discovery that SCFA are effective as a therapeuticapproach for the treatment and prevention of an allergic, autoimmuneand/or asthma disease or disorder.

In one embodiment, an autoimmune and/or allergic disease or disorder isat least one of Addison's disease, Agammaglobulinemia, Allergicrhinitis, Alopecia areata, Amyloidosis, Ankylosing spondylitis,Anti-GBM/Anti-TBM nephritis, Antiphospholipid syndrome (APS), Asthma,Autoimmune inner ear disease (AIED), Axonal & neuronal neuropathy(AMAN), Behcet's disease, Bullous pemphigoid, Castleman disease (CD),Celiac disease, Chagas disease, Chronic inflammatory demyelinatingpolyneuropathy (CIDP), Chronic recurrent multifocal osteomyelitis(CRMO), Churg-Strauss, Cicatricial pemphigoid/benign mucosal pemphigoid,Cogan's syndrome, Cold agglutinin disease, Congenital heart block,Coxsackie myocarditis, CREST syndrome, Crohn's disease, Dermatitisherpetiformis, Dermatomyositis, Devic's disease (neuromyelitis optica),Discoid lupus, Dressler's syndrome, Endometriosis, Eosinophilicesophagitis (EoE), Eosinophilic fasciitis, Erythema nodosum, Essentialmixed cryoglobulinemia, Evans syndrome, Fibromyalgia, Fibrosingalveolitis, Food allergies, Gastroenteritis, Giant cell arteritis(temporal arteritis), Giant cell myocarditis, Glomerulonephritis,Goodpasture's syndrome, Granulomatosis with Polyangiitis, Graves'disease, Guillain-Barre syndrome, Hashimoto's thyroiditis, Hemolyticanemia, Henoch-Schonlein purpura (HSP), Herpes gestationis or pemphigoidgestationis (PG), Hypogammalglobulinemia, IgA Nephropathy, IgG4-relatedsclerosing disease, Inclusion body myositis (IBM), Inflammatory boweldisease, Interstitial cystitis (IC), Juvenile arthritis, Juvenilerheumatoid arthritis, Juvenile diabetes (Type 1 diabetes), Juvenilemyositis (JM), Kawasaki disease, Lambert-Eaton syndrome,Leukocytoclastic vasculitis, Lichen planus, Lichen sclerosus, Ligneousconjunctivitis, Linear IgA disease (LAD), Lupus, Lyme disease chronic,Meniere's disease, Microscopic polyangiitis (MPA), Mixed connectivetissue disease (MCTD), Mooren's ulcer, Mucha-Habermann disease, Multiplesclerosis (MS), Myasthenia gravis, Myositis, Narcolepsy, Neuromyelitisoptica, Neutropenia, Ocular cicatricial pemphigoid, Optic neuritis,Palindromic rheumatism (PR), PANDAS (Pediatric AutoimmuneNeuropsychiatric Disorders Associated, with, Streptococcus),Paraneoplastic cerebellar degeneration (PCD), Paroxysmal nocturnalhemoglobinuria (PNH), Parry Romberg syndrome, Pars planitis (peripheraluveitis), Parsonnage-Turner syndrome, Pemphigus, Peripheral neuropathy,Perivenous encephalomyelitis, Pernicious anemia (PA), POEMS syndrome(polyneuropathy, organomegaly, endocrinopathy, monoclonal, gammopathy,skin changes), Polyarteritis nodosa, Polymyalgia rheumatica,Polymyositis, Postmyocardial infarction syndrome, Postpericardiotomysyndrome, Progesterone dermatitis, Psoriasis, Plaque psoriasis, Guttatepsoriasis, Inverse psoriasis, Pustular psoriasis, Psoriasis Vulgaris,Seborrheic Psoriasis, Erythrodermic psoriasis, Nail psoriasis, Psoriaticarthritis, Pure red cell aplasia (PRCA), Pyoderma gangrenosum, Raynaud'sphenomenon, Reactive Arthritis, Reflex sympathetic dystrophy, Reiter'ssyndrome, Relapsing polychondritis, Restless legs syndrome (RLS),Retroperitoneal fibrosis, Rheumatic fever, Rheumatoid arthritis (RA),Sarcoidosis, Schmidt syndrome, Scleritis, Scleroderma, Sjogren'ssyndrome, Sperm & testicular autoimmunity, Stiff person syndrome (SPS),Subacute bacterial endocarditis (SBE), Susac's syndrome, Sympatheticophthalmia (SO), Takayasu's arteritis, Temporal arteritis/Giant cellarteritis, Thrombocytopenic purpura (TTP), Tolosa-Hunt syndrome (THS),Transverse myelitis, Type 1 diabetes, Ulcerative colitis (UC),Undifferentiated connective tissue disease (UCTD), Uveitis, Vasculitis,Vitiligo, and Wegener's granulomatosis (Granulomatosis with Polyangiitis(GPA)).

In one embodiment, an allergic disease includes but is not limited tohay fever, food allergies, atopic dermatitis, allergic asthma,anaphylaxis, and the likes. Symptoms may include red eyes, an itchyrash, runny nose, shortness of breath, or inflammation. Exemplaryallergic diseases include, but are not limited to, Food allergies (e.g.,milk, soy, eggs, wheat, peanuts, tree nuts, fish, and shellfish), Latexallergies, Allergic rhinitis, Asthma, Atopic eczema, Anaphylaxis, Insectvenom, Drug allergies, and any combination thereof. The prevalence ofallergic diseases has increased all over the world during the last twodecades.

In one embodiment the invention is useful for the prevention of anautoimmune and/or allergic disease or disorder in a C-section deliveredneonate. Accordingly, the invention in one aspect encompassesadministration of SCFA to a subject, including expecting mothers,mothers, and neonates, as an effective way in reducing the risk ofdeveloping an autoimmune and/or allergic disease or disorder inneonates, and may also be effective in treating an autoimmune and/orallergic disease or disorder in neonates.

In one embodiment, the invention relates to compositions comprisingSCFAs for use as therapeutics for the treatment and prevention of anautoimmune and/or allergic disease or disorder in a C-section deliveredneonate. In one embodiment, the composition comprises one or more SCFAs.In one embodiment, the composition comprises one or more salts of one ormore SCFAs. In one embodiment, the composition comprises one or morebiologically active derivatives of one or more SCFAs. In one embodiment,the composition comprises one or more precursors of one or more SCFAs.In one embodiment, a composition comprises one or more combinations ofSCFAs, salts thereof, biologically active derivatives thereof, orprecursors thereof. In one embodiment, a composition comprises one ormore combinations of SCFAs, salts thereof, biologically activederivatives thereof, or precursors thereof, in combination with at leastone other compound. In one embodiment, a composition comprises one ormore of formic acid, acetic acid, propionic acid, isobutyric acid,butyric acid, tributyrin, N-acetylbutyrate (as well as other forms ofbutyrate), isovaleric acid, valeric acid, isocaproic acid, caproic acid,lactic acid, succinic acid, pyruvic acid, octanoic acid, and dodecanoicacid. In one embodiment, the composition is in the form of a capsule(enteric coated, time release) that can be used by women during latepregnancy. In another embodiment, the composition is in the form of abuffered solution of SCFAs that can be added to infant formula, pumpedbreast milk, baby food, and the like.

In one embodiment, the invention relates to a method of treating orpreventing an autoimmune and/or allergic disease or disorder in aC-section delivered neonate comprising administering a compositioncomprising a SCFA to one or more subjects. In one embodiment, theadministering a composition step occurs before birth of the neonate,wherein the composition is administered to the expectant mother. In oneembodiment, the expectant mother is near term (about month 8 or 9 ofpregnancy). In one embodiment, the SCFAs are given to the expectantmother orally for a time period of less than or equal to two months. Inone embodiment, the SCFAs are administered to the expectant mother inthe dose of 6 tablets per day for one week, followed by 3 tablets perday until the C-section is performed. In one embodiment, the dose of asingle tablet is 600 mg of one or more SCFAs or salts thereof. In oneembodiment, salts of one or more SCFAs are used, wherein the salts aresodium, magnesium, and/or calcium salts of one or more SCFAs. In oneembodiment, the administering a composition step occurs after birth ofthe neonate, wherein the composition is administered to the mother, theneonate, or both the mother and the neonate. In one embodiment, theSCFAs are administered to the neonate, in the form of a mixture withfood or drink. In one embodiment, the SCFAs are administered to theneonate, in the form of a supplement to baby formula or food for atleast the first 18-24 months of life.

In one embodiment, the present invention provides methods for treatment,inhibition, prevention, or reduction of an autoimmune and/or allergicdisease or disorder associated with C-section delivery of a neonate, byadministering a composition comprising a SCFA or a compound comprising aSCFA moiety, as disclosed herein, to a subject in need thereof,optionally in combination with at least one additional agent or therapy.In one embodiment, the subject is the expectant mother. In oneembodiment, the subject is the fetus. In one embodiment, the subject isthe mother. In one embodiment, the subject is the neonate. Autoimmuneand/or allergic diseases or disorders that may be treated using themethods of the invention include, but are not limited to any autoimmuneand/or allergic disease or disorder described elsewhere herein. In oneembodiment, the present invention provides methods for treatment,inhibition, prevention, or reduction of a disease associated with anautoimmune and/or allergic disease or disorder, comprising administeringa composition comprising a SCFA or a compound comprising a SCFA moiety,as disclosed herein, to a subject in need thereof.

In one embodiment, a subject at risk of developing, or having alreadydeveloped, an autoimmune or allergic disease or disorder is a human. Inone embodiment, a subject is a non-human animal.

In one embodiment, an exemplary method for the treatment or preventionof autoimmune and/or allergic diseases or disorder comprisesadministration of a daily oral dose of a composition comprising at least10 mg, at least 20 mg, at least 30 mg, at least 40 mg, at least 50 mg,at least 60 mg, at least 70 mg, at least 80 mg, at least 90 mg, at least100 mg, at least 200 mg, at least 300 mg, at least 400 mg, at least 500mg, at least 600 mg, at least 700 mg, at least 800 mg, at least 900 mg,at least 1 g, at least 2 g, at least 3 g, least 4 g, at least 5 g, atleast 6 g or more than 6 g of at least one SCFA at least 1 time daily,at least 2 times daily, at least 3 times daily or more than 3 timesdaily, for at least one week, at least two weeks, at least three weeks,at least one month, at least 2 months, at least 3 months, at least 4months, at least 5 months, at least 6 months or for more than 6 months.

In one embodiment, the composition comprising at least one SCFA is anenteric coated, extended release capsule.

In one embodiment, the composition may be administered, for example, toa breastfeeding mother of a C-section delivered neonate. In such anembodiment, an exemplary daily oral dose for healthy mothers is 1-2 g ofbutyrate and 0.5-1 g of propionate and acetate in enteric coated,extended release capsules, 3 times/day during first month, then half ofthat dose (e.g., 0.5-1 g of butyrate and 0.25-0.5 g of propionate andacetate) for at least 1, at least 2, at least 3 or more than 3additional months.

In one embodiment, the composition may be administered, for example, toa C-section delivered neonate. In such an embodiment, an exemplary dailyoral dose may be at least 10 mg, at least 20 mg, at least 30 mg, atleast 40 mg, at least 50 mg, at least 60 mg, at least 70 mg, at least 80mg, at least 90 mg, 100 mg, at least 200 mg, at least 300 mg, at least400 mg, at least 500 mg, at least 600 mg, at least 700 mg, at least 800mg, at least 900 mg, or more than 1 g of at least one SCFA at least 1time daily, at least 2 times daily, at least 3 times daily, at least 4times daily, at least 5 times daily, at least 6 times daily, at least 7times daily or more than 7 times daily. Such an embodiment may beformulated, for example as an additive to a nutritional formula, such asan infant food formula, for administration to a C-section deliveredneonate. In such an embodiment, an exemplary daily oral dose foradministration to a C-section delivered neonate comprises 80-100 mg ofbutyrate, 20-30 mg of acetate and propionate during the first monthfollowing delivery, followed by 100-120 mg of butyrate and 30-40 mg ofacetate and propionate for the subsequent 3-5 months.

Vasculitis

In one embodiment, the present invention provides methods for treatment,inhibition, prevention, or reduction of a vasculitis disease or disorderby administering a composition comprising a SCFA or a compoundcomprising a SCFA moiety, as disclosed herein, to a subject in needthereof, optionally in combination with at least one additional agent ortherapy. Vasculitis diseases and disorders that may be treated using themethods of the invention include, but are not limited to, lymphangitis,polymyalgia rheumatica, Takayasu's arteritis, temporal arteritis,Buerger's disease, Kawasaki disease, polyarteritis nodosa, Behçet'ssyndrome, eosinophilic granulomatosis with polyangiitis, cutaneousvasculitis, Henoch-Schönlein purpura, microscopic polyannulomatosis,cutaneous small-vessel vasculitis, granulomatosis with polyangiitis,Behçet's disease, and giant cell arteritis. In one embodiment, thepresent invention provides methods for treatment, inhibition,prevention, or reduction of a disease associated with vasculitis,comprising administering a composition comprising a SCFA or a compoundcomprising a SCFA moiety, as disclosed herein, to a subject in needthereof.

In various embodiments, one or more SCFA or compound comprising a SCFAmoiety may be used in combination with one or more corticosteroid drugs,such as prednisone or methylprednisolone (Medrol) for the treatment ofvasculitis. In other embodiments, one or more SCFA or compoundcomprising a SCFA moiety may be used in combination with one or morecytotoxic or immunosuppressant drugs, which decrease the function ofimmune system cells causing the inflammation. They include azathioprine(Azasan, Imuran), methotrexate (Trexall, Rheumatrex) andcyclophosphamide. In other embodiments, one or more SCFA or compoundcomprising a SCFA moiety may be used in combination with Rituximab(Rituxan), for treating some types of vasculitis.

In one embodiment, an exemplary method for the treatment of vasculitiscomprises administration of a daily oral dose of at least 100 mg, atleast 200 mg, at least 300 mg, at least 400 mg, at least 500 mg, atleast 600 mg, at least 700 mg, at least 800 mg, at least 900 mg, atleast 1 g, at least 2 g, at least 3 g, at least 4 g, at least 5 g, atleast 6 g or more than 6 g of at least one SCFA, at least 1 time daily,at least 2 times daily, at least 3 times daily or more than 3 timesdaily, for at least one week, at least two weeks, at least three weeks,at least one month, at least 2 months, at least 3 months, at least 4months, at least 5 months, at least 6 months or for more than 6 months.

In one embodiment, the composition comprising at least one SCFA is anenteric coated, extended release capsule.

In one embodiment, an exemplary method for the treatment of vasculitiscomprises administration of a daily oral dose of 5 g-6 g of butyrate inenteric coated, extended release capsules, at least 3 times daily forone month, followed administration of a daily oral dose of 3 g-4 g ofbutyrate in enteric coated, extended release capsules, at least 2 timesdaily for at least 2 months. In one embodiment, higher doses can be usedfor the treatment of disease flares, including, but not limited to,stress triggered disease flares.

In one embodiment, the method includes administering a compositioncomprising a SCFA in combination with other steroids and/orchemotherapeutic agents. In such an embodiment, the dosage level of theother steroids and/or chemotherapeutic agents may be reduced relative tothe recommended or administered dosage for a subject undergoingtreatment with the steroids and/or chemotherapeutic agents alone.

Lymphoma

In one embodiment, the present invention provides methods for treatment,inhibition, prevention, or reduction of a lymphoma disease or disorderby administering a composition comprising a SCFA or a compoundcomprising a SCFA moiety, as disclosed herein, to a subject in needthereof, optionally in combination with at least one additional agent ortherapy. Lymphoma diseases and disorders that may be treated using themethods of the invention include, but are not limited to activatedB-cell-like diffuse large B cell lymphoma (ABC DLBCL), follicularlymphoma (FL), mucosa associated lymphoid tissue lymphoma (MALT),Hodgkin lymphoma (HL), and primary mediastinal B cell lymphoma (PMBL).In one embodiment, the present invention provides methods for treatment,inhibition, prevention, or reduction of a disease associated withlymphoma, or prevention of tumor relapse, comprising administering acomposition comprising a SCFA or a compound comprising a SCFA moiety, asdisclosed herein, to a subject in need thereof.

In various embodiments, one or more SCFA or compound comprising a SCFAmoiety may be used in combination with one or more other lymphomatreatment therapies. In various embodiments, the one or more otherlymphoma treatment therapies includes radiation therapy, and/orrituximab (anti-CD20 antibody). In various embodiments, a composition ofthe invention is administered before, during, or after another treatmentfor lymphoma. In various embodiments, another treatment for lymphomaincludes one or more chemotherapeutic agents, anti-proliferative agents,anti-tumor agents, antineoplastic agents, anti-angiogenic agents, and/orother anti-cancer agents as disclosed elsewhere herein.

In one embodiment, the method includes administering a compositioncomprising a SCFA in combination with other anti-lymphoma or anti-tumoragents, including chemotherapeutic agents, cytotoxic/anti-neoplasticagents and anti-angiogenic agents. In such an embodiment, the dosagelevel of the other anti-lymphoma or anti-tumor agents may be reducedrelative to the recommended or administered dosage for a subjectundergoing treatment with the other anti-lymphoma or anti-tumor agentsalone.

Leukemia

In one embodiment, the present invention provides methods for treatment,inhibition, prevention, or reduction of a leukemia disease or disorderby administering a composition comprising a SCFA or a compoundcomprising a SCFA moiety, as disclosed herein, to a subject in needthereof, optionally in combination with at least one additional agent ortherapy. Leukemia diseases and disorders that may be treated using themethods of the invention include, but are not limited tomyeloproliferative neoplasm (MPN), polycythemia vera (PV), essentialthrombocytosis (ET), idiopathic/myelofibrosis (Mf), acute myeloidleukemia (AML), childhood acute lymphoblastic leukemia (ALL), and bloodcancer. In one embodiment, the present invention provides methods fortreatment, inhibition, prevention, or reduction of a disease associatedwith leukemia, for treating or preventing relapse, or for prevention ofprimary tumor onset, of at least one type of leukemia, or for preventingthe evolution of MPN to leukemia comprising administering a compositioncomprising a SCFA or a compound comprising a SCFA moiety, as disclosedherein, to a subject in need thereof. In one embodiment, the at leastone SCFA or compound comprising a SCFA moiety is administered before,during, or after the administration of at least one additional agent ortherapy for the treatment or prevention of leukemia. In a particularembodiment, a composition of the invention is administered after atherapeutic phase as part of a strategy to prevent relapse, or forprevention of primary tumor onset, of the leukemia. In variousembodiments, a composition of the invention is administered before,during, or after another treatment for leukemia. In various embodiments,another treatment for leukemia includes one or more chemotherapeuticagents, antiproliferative agents, anti-tumor agents, antineoplasticagents, antiangiogenic agents, and/or other anti-cancer agents asdisclosed elsewhere herein.

In one embodiment, treatment comprises two phases. The first phase,referred to as induction, may include chemotherapy, for example,treatment with arabinosylcytosine (ara-C) and daunomycin, optionallycombined with, cladribine (Leustatin, 2-CdA). Induction is consideredsuccessful if remission is achieved. The second phase, referred to asconsolidation, includes long term high dose ara-C. Alternatively,allogeneic or autologous stem cell transplant may be used. In oneembodiment, a SCFA therapeutic compound of the invention is administeredto the subject before, during, and/or after induction. In anotherembodiment, a SCFA therapeutic compound of the invention is administeredto the subject before, during, and/or after consolidation. In oneembodiment, a SCFA therapeutic compound of the invention is administeredto the subject before, during, and/or after induction and before,during, and/or after consolidation.

In various embodiments, a composition of the invention is administeredas part of a method to treat or prevent relapse, or for prevention ofprimary tumor onset, of at least one leukemia. In one embodiment, the atleast one leukemia depends upon constitutive activation of JAK/STATsignaling. In one embodiment, the at least one leukemia is BCR/ABLnegative. In one embodiment, the treatment method delays or preventsleukemia recurrence or relapse, or for prevention of primary tumoronset. In one embodiment, the treatment method decreases the toxicityprofiles of standard of care treatments started at the time ofdiagnosis, thereby increasing long term quality of life.

In one embodiment, the method includes administering a compositioncomprising a SCFA in combination with other anti-leukemia or anti-tumoragents, including chemotherapeutic agents, cytotoxic/anti-neoplasticagents and anti-angiogenic agents. In such an embodiment, the dosagelevel of the other anti-leukemia or anti-tumor agents may be reducedrelative to the recommended or administered dosage for a subjectundergoing treatment with the other anti-leukemia or anti-tumor agentsalone.

Immunotherapy

In one embodiment, the invention provides a method for stimulating,eliciting or enhancing an immune response in a subject individual(individual) comprising administering to a subject an immunotherapeuticagent in combination with a composition comprising a SCFA or a compoundcomprising a SCFA moiety. In one embodiment, the subject may be at riskfor having a disease, be diagnosed as having a disease, have previouslybeen treated for a disease, or be contemporaneously be treated for thedisease using a treatment method (e.g., a treatment method that does notinclude the use of a composition of the invention described herein).

The present invention is partly based upon the discovery thatadministration of at least one short chain fatty acid (SCFA) can beeffective as a therapeutic approach for the treatment or prevention ofan adverse effect associated with immunotherapy of cancer. It isexpected that the therapeutic effect of treatments using compositionscomprising at least one SCFA will not interfere significantly with thebenefits of the immunotherapy, but will treat or prevent undesiredadverse effects associated with immunotherapy of cancer. Therefore, inone embodiment, the invention relates to compositions comprising atleast one SCFA for use as a therapeutic for the treatment or preventionof adverse effects associated with immunotherapy of cancer.

In one embodiment, the present invention provides methods for treatment,inhibition, prevention, or reduction of an adverse effect associatedwith cancer immunotherapy by administering a composition comprising aSCFA or a compound comprising a SCFA moiety, as disclosed herein, to asubject in need thereof, optionally in combination with at least oneadditional agent or therapy. Adverse effects associated with cancerimmunotherapy that may be treated using the methods of the inventioninclude, but are not limited to cytokine release syndrome (CRS),neurological toxicity, on-target/off-tumor recognition, anaphylaxis,graft versus host disease (GVHD), off-target antigen recognition, andmacrophage activation syndrome (MAS). In one embodiment, the presentinvention provides methods for treatment, inhibition, prevention, orreduction of a disease associated with an adverse effect associated withcancer immunotherapy, comprising administering a composition comprisingat least one SCFA or compound comprising a SCFA moiety, as disclosedherein, to a subject in need thereof. In one embodiment, the adverseeffect includes a combination of adverse effects.

In one embodiment, a subject suffering from adverse effects associatedwith immunotherapy of cancer is a human. In one embodiment, a subjectsuffering from adverse effects associated with immunotherapy of canceris a non-human animal.

In one embodiment, to stimulate an immune response, the subjectindividual is administered (i) at least one SCFA or compound comprisinga SCFA moiety, and (ii) at least one immunotherapeutic agent. Typically,the administration of at least one SCFA or compound comprising a SCFAmoiety and the immunotherapeutic agent will be in the form of a vaccineor administered in a vaccine regimen. The at least one SCFA or compoundcomprising a SCFA moiety and the immunotherapeutic agent can beadministered at about the same time to the subject individual, or can beadministered separately and/or sequentially.

Immunotherapeutic agents that may be administered according to themethods of the invention include, but are not limited to, one or morecancer antigens, one or more antigens derived from a virus associatedwith cancer, and an anti-cancer antibody.

A cancer antigen may be (a) a cell surface antigen that can be found ona malignant cell, (b) an antigen that can be found inside a malignantcell or (c) a mediator of tumor cell growth. The term “cancer antigen”refers to (i) tumor-specific antigens, (ii) tumor-associated antigens,(iii) cells that express tumor-specific antigens, (iv) cells thatexpress tumor-associated antigens, (v) embryonic antigens on tumors,(vi) autologous tumor cells, (vii) tumor-specific membrane antigens,(viii) tumor-associated membrane antigens, (ix) growth factor receptors,(x) growth factor ligands, and (xi) any other type of antigen orantigen-presenting cell or material that is associated with a cancer.

The cancer antigen can be a cell, a protein, a peptide, a fusionprotein, DNA encoding a peptide or protein, RNA encoding a peptide orprotein, a glycoprotein, a lipoprotein, a phosphoprotein, acarbohydrate, a lipopolysaccharide, a lipid, a chemically linkedcombination of two or more thereof, a fusion or two or more thereof, ora mixture of two or more thereof. In another embodiment, the cancerantigen is a peptide comprising about 6 to about 24 amino acids; fromabout 8 to about 20 amino acids; from about 8 to about 12 amino acids;from about 8 to about 10 amino acids; or from about 12 to about 20 aminoacids. In one embodiment, the cancer antigen is a peptide having a MEWClass I binding motif or a MHC Class II binding motif. In anotherembodiment, the cancer antigen comprises a peptide that corresponds toone or more cytotoxic T lymphocyte (CTL) epitopes.

In one embodiment, the present invention provides methods for treatment,inhibition, prevention, or reduction of an adverse effect associatedwith cancer immunotherapy by administering a composition comprising aSCFA or a compound comprising a SCFA moiety, as disclosed herein, to asubject in need thereof, optionally in combination with at least oneimmunotherapeutic agent.

In one embodiment, the immunotherapy is chimeric antigen receptor T-cell(CAR-T) therapy. In some embodiments, a composition of the invention isadministered before, during, or after CAR-T therapy for the treatment ofcancer, for the treatment or prevention of at least one adverse effectassociated with CAR-T therapy. Particular adverse effects treatableand/or preventable by compositions of the present invention include, butare not limited to: cytokine release syndrome (CRS), neurologicaltoxicity, on-target/off-tumor recognition, anaphylaxis, graft versushost disease (GVHD), off-target antigen recognition, and macrophageactivation syndrome (MAS). Existing or developing treatment orprevention of these adverse effects include pharmacologicalimmunosuppression (i.e., IL-6R blockade, systemic corticosteroids suchas dexamethasone, monoclonal antibodies, lymphodepleting chemotherapywith agents such as cyclophosphamide), suicide genes or eliminationgenes (i.e., killing of CAR-T cells), and targeted activation (i.e.,conditional activation of CAR-T cells with a drug or another agent).Therefore, in one embodiment, the treatment compositions of the presentinvention are combined with at least one other strategy to reduce,prevent, treat, or ameliorate one or more adverse effects associatedwith immunotherapy of cancer. In one embodiment, the invention providesa composition for treating or preventing the onset of an adverse effectassociated with immunotherapy of cancer. In one embodiment, thecomposition comprises at least one SCFA or a compound comprising a SCFAmoiety. In one embodiment, the immunotherapy comprises CAR-T therapy. Inanother embodiment, the immunotherapy comprises a therapy involving atleast one other anti-cancer composition. In some embodiments, acomposition of the invention is administered before, during, or afterchemotherapy for the treatment of cancer.

In one embodiment, the composition comprising at least one SCFA isadministered in parallel to chemotherapy: several days before CAR-T cellinfusion (only SCFAs), and up to several weeks after CAR-T cell infusion(SCFAs in combination with chemotherapeutic agents at reduced levelsrelative to a subject undergoing chemotherapy alone).

In one embodiment, an exemplary daily oral dose for use in methods ofimmunotherapy is daily oral dose of at least 2 g, at least 3 g, at least4 g, at least 5 g, at least 6 g or more than 6 g of at least one SCFA,at least 2 times daily, at least 3 times daily or more than 3 timesdaily, for at least one week, at least two weeks, at least three weeks,at least one month, at least 2 months, at least 3 months, at least 4months, at least 5 months, at least 6 months or for more than 6 months.

In one embodiment, an exemplary daily oral dose for use in methods ofimmunotherapy is daily oral dose of 5-6 g of butyrate and 2-3 g ofpropionate. In one embodiment, the composition comprising at least oneSCFA is an enteric coated, extended release capsule. In one embodiment,the composition is administered 3 times/day.

Combination with Additional Agents

The present invention is also directed to a method of treating orpreventing a disease or disorder as described above in combination withone or more additional agents.

The combination can be in a single formulation or can be separate andadministered in sequence (either a composition comprising at least oneSCFA, or a molecule comprising a SCFA moiety, first and then acomposition comprising an additional agent, or a composition comprisingan additional agent first and then a composition comprising at least oneSCFA, or a molecule comprising a SCFA moiety). In some embodiments, theat least one SCFA, or a molecule comprising a SCFA moiety, can beadministered to the subject about 30 seconds, 1 minute, 2 minutes, 3minutes, 4 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 25minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55minutes, 60 minutes, 0.25 hours, 0.5 hours, 0.75 hours, 1 hours, 2hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24hours, 36 hours, 48 hours, 60 hours, 72 hours, 84 hours, 96 hours, 1day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26days, 27 days, 28 days, 29 days, 30 days, 31 days, 1 week, 2 weeks, 3weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks before thecomposition comprising at least one additional agent is administered tothe subject. In other embodiments, the composition comprising at leastone additional agent can be administered to the subject about 30seconds, 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 10minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40minutes, 45 minutes, 50 minutes, 55 minutes, 60 minutes, 0.25 hours, 0.5hours, 0.75 hours, 1 hours, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours,7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21hours, 22 hours, 23 hours, 24 hours, 36 hours, 48 hours, 60 hours, 72hours, 84 hours, 96 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30days, 31 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7weeks, or 8 weeks before the composition comprising at least one SCFA,or a molecule comprising a SCFA moiety, is administered to the subject.

In one embodiment, the invention provides a general concept foradministering at least one of these agents, or a biologically-activederivative thereof, in combination with at least one short chain fattyacid (SCFA), or a biologically-active derivative or precursor thereof,as a therapy for treating or preventing a disease or disorder in asubject in need thereof. In one embodiment, the composition of theinvention comprises at least one of these agents, or abiologically-active derivative thereof, and at least one SCFA, or abiologically-active derivative or precursor thereof.

Exemplary therapeutic agents that can be included in a composition ofthe invention or administered in combination with the composition of theinvention are provided.

Checkpoint Inhibitors

In one embodiment, the method comprises administering to a subject inneed thereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, in combination withone or more immune checkpoint inhibitor. “Checkpoint inhibitor” as usedherein includes inhibitors or molecules that block immune checkpoints ascommonly understood in the field of cancer immunotherapy. Commonly thecheckpoint inhibitors are antibodies that block the immune checkpointproteins. Immune checkpoint proteins include, but are not limited to,PD1, PDL1, PDL2, CTLA-4, LAG3, TIM3, B7-H3, BTLA, VISTA, CD40, CEACAM1,CD80, CD86, OX40, CD27, GITR, DNAM-1, TIGIT, TMIGD2 and DC-SIGN. Someexamples of known checkpoint inhibitors include, but are not limited to,ipilimumab, pembrolizumab, nivolumab, pidilizumab, atezolizumab,avelumab, druvbalumab and others. In one embodiment, the composition asdescribed above can comprise at least one SCFA, or a molecule comprisinga SCFA moiety, in combination with an antibody to a checkpoint protein.

In one embodiment, the combination of the at least one SCFA, or amolecule comprising a SCFA moiety, and checkpoint inhibitor induces theimmune system more efficiently than the checkpoint inhibitor alone. Thismore efficient immune response provides increased efficacy in thetreatment and/or prevention of a particular disease or disorder.

In some embodiments, the immune response can be increased by about0.5-fold to about 15-fold, about 0.5-fold to about 10-fold, or about0.5-fold to about 8-fold. Alternatively, the immune response in thesubject administered the combination of the at least one SCFA, or amolecule comprising a SCFA moiety, and checkpoint inhibitor can beincreased by at least about 0.5-fold, at least about 1.0-fold, at leastabout 1.5-fold, at least about 2.0-fold, at least about 2.5-fold, atleast about 3.0-fold, at least about 3.5-fold, at least about 4.0-fold,at least about 4.5-fold, at least about 5.0-fold, at least about5.5-fold, at least about 6.0-fold, at least about 6.5-fold, at leastabout 7.0-fold, at least about 7.5-fold, at least about 8.0-fold, atleast about 8.5-fold, at least about 9.0-fold, at least about 9.5-fold,at least about 10.0-fold, at least about 10.5-fold, at least about11.0-fold, at least about 11.5-fold, at least about 12.0-fold, at leastabout 12.5-fold, at least about 13.0-fold, at least about 13.5-fold, atleast about 14.0-fold, at least about 14.5-fold, or at least about15.0-fold.

In still other alternative embodiments, the immune response in thesubject administered the combination of the at least one SCFA, or amolecule comprising a SCFA moiety, and checkpoint inhibitor can beincreased about 50% to about 1500%, about 50% to about 1000%, or about50% to about 800%. In other embodiments, the immune response in thesubject administered the combination of the at least one SCFA, or amolecule comprising a SCFA moiety, and checkpoint inhibitor can beincreased by at least about 50%, at least about 100%, at least about150%, at least about 200%, at least about 250%, at least about 300%, atleast about 350%, at least about 400%, at least about 450%, at leastabout 500%, at least about 550%, at least about 600%, at least about650%, at least about 700%, at least about 750%, at least about 800%, atleast about 850%, at least about 900%, at least about 950%, at leastabout 1000%, at least about 1050%, at least about 1100%, at least about1150%, at least about 1200%, at least about 1250%, at least about 1300%,at least about 1350%, at least about 1450%, or at least about 1500%.

Anti-Inflammatory Agents

In one embodiment, the method comprises administering to a subject inneed thereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, in combination withone or more anti-inflammatory agent. Exemplary anti-inflammatory agentsthat can be used in combination with the compositions of the inventioninclude, but are not limited to nonsteroidal anti-inflammatory drugs(NSAIDs) such as diclofenac (e.g., Arthrotec®), diflunisal (e.g.,Dolobid®), etodolac (e.g., Lodine®), fenoprofen (e.g., Nalfon®),ibuprofen (e.g., Advil®, Motrin®, and others), indomethacin (e.g.,Arthrexin®), ketoprofen (e.g., Oruvail®) ketorolac (e.g., Toradol®),fosfomycin tromethamine (e.g., Monural®), meclofenamate (e.g.,Meclomen®), nabumetone (e.g., Relafen®), naproxen (e.g., Anaprox®, andothers). Oxaprozin (e.g., Daypro®) piroxicam (e.g., Feldene®) sulindac(e.g., tolmetin (e.g., Tolectin®, and others), Flavenoids (e.g.,luteolin, fisetin, and apigenin), steroids, antihistamines, Loratadine,Theophylline, Doxantrazole, Quercetin, 8-bromo cyclic AMP, Disodiumcromoglicate, Beclomethasone dipropionate, Budesonide,Budesonide/Formoterol, Fluticasone, Fluticasone inhaled powder,Fluticasone/Salmeterol, Mometasone, Mometasone/formoterol, Cromolyn,Omalizumab, Inhaled short- or long-acting beta2-agonists, Leukotrienemodifiers, and Theophylline. In one embodiment, the method comprisesadministering to a subject in need thereof a composition comprising atleast one SCFA, or a biologically-active derivative or precursorthereof, and additionally one or more anti-inflammatory agent. In oneembodiment, the method comprises administering to a subject in needthereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, in combination witha composition comprising one or more anti-inflammatory agent.

PDE4 Inhibitors

In one embodiment, the method comprises administering to a subject inneed thereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, in combination withone or more phosphodiesterase 4 (PDE4) inhibitors. A non-limitingselection of PDE4 inhibitors is presented in U.S. Patent ApplicationPublication No. 2002/0111495, which is incorporated herein by reference.In one embodiment, the method comprises administering to a subject inneed thereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, and additionallyone or more PDE4 inhibitor. In one embodiment, the method comprisesadministering to a subject in need thereof a composition comprising atleast one SCFA, or a biologically-active derivative or precursorthereof, in combination with a composition comprising one or more PDE4inhibitor.

Disease-Modifying Antirheumatic Drugs (DMARD)

In one embodiment, the method comprises administering to a subject inneed thereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, in combination withone or more disease-modifying antirheumatic drugs (DMARD) orimmunosuppressants may be included in the compositions of the invention.DMARDs are known in the art, and include, but are not limited to,methotrexate (Rheumatrex®), sulfasalazine (Azulfidine®), mycophenolatemofetil (CellCept®), and cyclosporine (Sandimmune®, or Neroal®). In oneembodiment, the method comprises administering to a subject in needthereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, and additionallyone or more DMARD. In one embodiment, the method comprises administeringto a subject in need thereof a composition comprising at least one SCFA,or a biologically-active derivative or precursor thereof, in combinationwith a composition comprising one or more DMARD.

Biologic Agents

In one embodiment, the method comprises administering to a subject inneed thereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, in combination withone or more biologic drugs. Exemplary biologic drugs contemplated by thepresent invention include, but are not limited to, etanercept (Enbrel®),infliximab (Remicade®), apremilast (Otezla®), and adalimumab (Humira®).In one embodiment, the method comprises administering to a subject inneed thereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, and additionallyone or more biologic agent. In one embodiment, the method comprisesadministering to a subject in need thereof a composition comprising atleast one SCFA, or a biologically-active derivative or precursorthereof, in combination with a composition comprising one or morebiologic agent.

Cox-2 Inhibitors

In one embodiment, the method comprises administering to a subject inneed thereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, in combination withone or more Cox-2 inhibitors. Cox-2 inhibitors include, but are notlimited to, celecoxib (Celebrex®), valdecoxib (Bextra®), and meloxicam(Mobic®). In one embodiment, the method comprises administering to asubject in need thereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, and additionallyone or more Cox-2 inhibitors. In one embodiment, the method comprisesadministering to a subject in need thereof a composition comprising atleast one SCFA, or a biologically-active derivative or precursorthereof, in combination with a composition comprising one or more Cox-2inhibitors.

Magnesium, Vitamin D3, and Vitamin E

In one embodiment, the method comprises administering to a subject inneed thereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, and additionallyone or more of magnesium, vitamin D3, and vitamin E (d-α-tocopherolacetate). Magnesium is a co-factor for more than 300 enzymes thatregulate diverse biochemical reactions including regulation of bloodglucose levels, detoxification, and others. Vitamin D3 deficiency isfrequent in patients with immune disorders. Vitamin E has distinctiveantioxidant activities. In one embodiment, the method comprisesadministering to a subject in need thereof a composition comprising atleast one SCFA, or a biologically-active derivative or precursorthereof, and additionally one or more of magnesium, vitamin D3, andvitamin E. In one embodiment, the method comprises administering to asubject in need thereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, in combination witha composition comprising one or more of magnesium, vitamin D3, andvitamin E.

Other Agents

In one embodiment, the method comprises administering to a subject inneed thereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, and additionallyone or more additional therapeutic agents. In one embodiment, the methodcomprises administering to a subject in need thereof a compositioncomprising at least one SCFA, or a biologically-active derivative orprecursor thereof, in combination with a composition comprising one ormore additional therapeutic agents. Additional therapeutic agents thatare contemplated for administration according to the methods of theinvention include, but are not limited to, rofecoxib, celecoxib, folicacid, sulfasalazine, naproxen, leflunomide, methylprednisolone acetate,parenteral gold, oral gold, indomethacin, hydroxychloroquine,hydroxychloroquine sulfate, sulindac, prednisone, betamethasone dipropaugmented, folate, triamcinolone acetonide, diclofenac,dimethylsulfoxide, piroxicam, diclofenac sodium, ketoprofen, meloxicam,methylprednisolone, nabumetone, tolmetin sodium, calcipotriene,cyclosporine, diclofenac, sodium/misoprostol, fluocinonide, glucosaminesulfate, gold sodium thiomalate, hydrocodone bitartrate/apap,risedronate sodium, sulfadiazine, thioguanine, valdecoxib, smallmolecule inhibitor of KDR (ABT-123), small molecule inhibitor of Tie-2,clobetasol propionate, triamcinolone acetonide, halobetasol propionate,tazarotene, fluocinonide, betamethasone diprop augmented, fluocinolone,acetonide, acitretin, tar shampoo, betamethasone valerate, mometasonefuroate, ketoconazole, pramoxine/fluocinolone, hydrocortisone valerate,flurandrenolide, urea, betamethasone, clobetasol propionate/emoll,fluticasone propionate, azithromycin, hydrocortisone, moisturizingformula, folic acid, desonide, coal tar, diflorasone diacetate, folate,lactic acid, methoxsalen, methylprednisolone acetate, prednisone,sunscreen, salicylic acid, halcinonide, anthralin, clocortolonepivalate, coal extract, coal tar/salicylic acid, coal tar/salicylicacid/sulfur, desoximetasone, diazepam, emollient, pimecrolimusemollient, fluocinonide/emollient, mineral oil/castor oil/na lact,mineral oil/peanut oil, petroleum/isopropyl myristate, psoralen,salicylic acid, soap/tribromsalan, thimerosal/boric acid, celecoxib,alefacept, efalizumab, tacrolimus, pimecrolimus, PUVA, UVB,sulfasalazine, alemtuzumab, dronabinol, Unimed, daclizumab,mitoxantrone, xaliproden hydrochloride, fampridine, glatiramer acetate,natalizumab, sinnabidol, a-immunokine NNSO3, ABR-215062, AnergiX.MS,chemokine receptor antagonists, BBR-2778, calagualine, CPI-1189, LEM(liposome encapsulated mitoxantrone), THC.CBD (cannabinoid agonist)MBP-8298, mesopram (PDE4 inhibitor), MNA-715, anti-IL-6 receptorantibody, neurovax, pirfenidone allotrap 1258 (RDP-1258), sTNF-R1,CDP571 (a humanized monoclonal anti-TNF-alpha IgG4 antibody), CDP 870 (ahumanized monoclonal anti-TNF-alpha antibody fragment), an anti-TNF dAb(Peptech), CNTO 148 (golimumab; Medarex and Centocor, see WO 02/12502),and adalimumab (Humira® Abbott Laboratories, a human anti-TNF mAb,described in U.S. Pat. No. 6,090,382 as D2E7). Additional TNF antibodieswhich can be used in the invention are described in U.S. Pat. Nos.6,593,458; 6,498,237; 6,451,983; and 6,448,380, each of which isincorporated by reference herein, talampanel, teriflunomide, TGF-beta2,tiplimotide, VLA-4 antagonists (for example, TR-14035, VLA4 Ultrahaler,Antegran-ELAN/Biogen), interferon gamma antagonists, IL-4 agonists,humanized IL-6 antibody tocilizumab, steroids (e.g., dexamethasone,prednisone, prednisolone, triamcinolone acetonide, fluorometholone, anddifluprednate), rapamycin, lampalizumab, fluocinolone acetatonide,macuCLEAR eyedrops, bone marrow CD34 stem cells, other stem cells,ranibizumab, brimonidine, LFG316, ORACEA®, emixustat hydrochloride,sirolimus, copaxone, othera eye drops, AL-78898A, and eculizumab.

Interleukin IL-17 Inhibitors

Interleukin IL-17 has been shown to stimulate TNFα production.Anti-IL-17 drugs include, but are not limited to brodalumab, ixekizumaband secukinumab. In one embodiment, the method comprises administeringto a subject in need thereof a composition comprising at least one SCFA,or a biologically-active derivative or precursor thereof, andadditionally one or more inhibitors of IL-17. In one embodiment, themethod comprises administering to a subject in need thereof acomposition comprising at least one SCFA, or a biologically-activederivative or precursor thereof, in combination with a compositioncomprising one or more inhibitors of IL-17.

IFN-γ Inhibitors

A key Th1-type cytokine involved in the pathogenesis of inflammatorydisease is IFN-γ which inhibits apoptosis of keratinocytes bystimulating expression of the anti-apoptotic protein Bcl-x. Thiscontributes to the progression of inflammatory disease. Therefore, inone embodiment, the method of the invention may comprise administrationof the composition of the invention in combination with one or moreinhibitors of IFN-γ. In one embodiment, an inhibitor of IFN-γ may be atleast one of an anti-IFN-γ antibody, a soluble IFN-γ receptor, and acombination thereof. In one embodiment, the method comprisesadministering to a subject in need thereof a composition comprising atleast one SCFA, or a biologically-active derivative or precursorthereof, and additionally one or more inhibitors of IFN-γ. In oneembodiment, the method comprises administering to a subject in needthereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, in combination witha composition comprising one or more inhibitors of IFN-γ.

VEGF Inhibitors

Vascular endothelial growth factor (VEGF) is implicated in thepathologic angiogenesis observed in inflammatory disease which ischaracterized by enhanced expression of VEGF (by epidermalkeratinocytes) at mRNA and protein levels, and VEGF receptors (bytortuous microvessels in the upper dermis) (Man et al., 2008, J Cell MolMed., 12(2):649-60; Marina et al., 2015, Clujul Med, 88(3):247-52).Therefore, in one embodiment, the method of the invention may compriseadministration of the composition of the invention in combination withat least one VEGF inhibitor. In one embodiment, the VEGF inhibitor is anantibody, for example, a monoclonal antibody. In one embodiment, theVEGF inhibitor is at least one of bevacizumab, sunitinib, sorafenib,pazopanib, brivanib alaninate, cediranib, vandetanib, motesanib,linifinib, axitinib, and aflibercept. In one embodiment, the methodcomprises administering to a subject in need thereof a compositioncomprising at least one SCFA, or a biologically-active derivative orprecursor thereof, and additionally one or more VEGF inhibitor. In oneembodiment, the method comprises administering to a subject in needthereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, in combination witha composition comprising one or more VEGF inhibitor.

HDAC Inhibitors

Butyrate is one of the strongest histone deacetylase (HDAC) inhibitors.HDAC inhibition may stimulate differentiation of naïve T-cells into Tregcells, a subset of T_(CD4) cells (expressing transcription factor FoxP3,which is crucial for Treg development). Treg cells are involved in themaintenance of self-tolerance by suppressing activation and expansion ofself-reactive lymphocytes and thus have a critical role in preventingautoimmunity. Therefore, the present invention provides a number ofcompositions that could promote Tregs and/or cells and/or pathwaysassociated with FoxP3, at least in part by HDAC inhibition. Therefore,in one embodiment, the composition of the invention comprises at leastone compound that promotes the differentiation of naïve T-cells intoTreg cells.

Therefore, in one embodiment, the method of the invention may compriseadministration of the composition of the invention in combination withone or more HDAC inhibitor. Exemplary HDAC inhibitor include, but arenot limited to, TSA, SAHA, CBHA, LAQ-824, PDX-101, LBH-589, ITF2357,PCI-24781, FK-228, valproic acid, phenyl butyrate, butyrate, AN-9,MS-275, and MGCD0103 (Dokmanovic, M. et al., 2007, 5(10):981-989). Inone embodiment, the method comprises administering to a subject in needthereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, and additionallyone or more HDAC inhibitor. In one embodiment, the method comprisesadministering to a subject in need thereof a composition comprising atleast one SCFA, or a biologically-active derivative or precursorthereof, in combination with a composition comprising one or more HDACinhibitor.

AMPK Activators

Adenosine monophosphate kinase (AMPK) promotes expression oftranscription factor FoxP3, which in turn stimulates Treg celldifferentiation and function. In one embodiment, the method of theinvention may comprise administration of the composition of theinvention in combination with one or more natural AMPK activators,including but not limited to, rosehip (from Rosa canina) and jiaogulanextracts (from Gynostemma pentaphyllum) or other natural AMPKactivators. Exemplary AMPK activators from natural products, aredescribed in Uddin et al., 2013, Natural Product Sciences, 19(1):1-7,which is incorporated herein by reference.

IDO Inhibitors

In one embodiment, the method of the invention may compriseadministration of the composition of the invention in combination withone or more IDO inhibitor. IDO inhibitors including, but are not limitedto, 1-methyl-DL-tryptophan; p-(3-benzofuranyl)-DL-alanine;p-[3-benzo(b)thienyl]-DL-alanine; and 6-nitro-L-tryptophan have beenused to modulate T cell-mediated immunity by altering localextracellular concentrations of tryptophan and tryptophan metabolites(WO 99/29310). Compounds having IDO inhibitory activity are furtherreported in WO 2004/094409, both the contents of which are incorporatedin their entirety herein.

In one embodiment, the method comprises administering to a subject inneed thereof a composition comprising at least one SCFA, or abiologically-active derivative or precursor thereof, and additionallyone or more IDO inhibitor. In one embodiment, the method comprisesadministering to a subject in need thereof a composition comprising atleast one SCFA, or a biologically-active derivative or precursorthereof, in combination with a composition comprising one or more IDOinhibitor.

Agents for the Treatment of Diabetes or Diabetes Related Conditions

In one embodiment, the method of the invention may compriseadministration of the composition of the invention in combination withone or more agent for the treatment of diabetes or diabetes relatedconditions. The use of other agents (e.g., insulin) and diets (lowsugar) can be used in combination with the SCFA or compound comprising aSCFA moiety of the present invention to increase therapeutic efficacythereof. Anti-diabetes compounds that may be combined with the SCFA orcompound comprising a SCFA moiety of the invention include, but are notlimited to, Short-acting insulin (e.g., regular insulin), Rapid-actinginsulins (e.g., insulin aspart, insulin glulisine, and insulin lispro),Intermediate-acting insulin (e.g., insulin isophane), Long-actinginsulins (e.g., insulin degludec, insulin detemir, insulin glargine, andinsulin glargine,), Combination insulins (e.g., 70/30 (insulin aspartprotamine-insulin aspart), 75/25 (insulin lispro protamine-insulinlispro), 50/50 (insulin lispro protamine-insulin lispro), 70/30 (humaninsulin NPH-human insulin regular), 70/30 (human insulin NPH-humaninsulin regular), and (insulin degludec-insulin aspart)), andPramlintide, an amylinomimetic drug.

Chemotherapeutic Agents

In one embodiment, the method of the invention may compriseadministration of the composition of the invention in combination withone or more chemotherapeutic agent. Chemotherapeutic agents include, butare not limited to, ara-C, daunomycin, cladribine (Leustatin, 2-CdA),cytotoxic agents (e.g., 5-fluorouracil, cisplatin, carboplatin,methotrexate, daunorubicin, doxorubicin, vincristine, vinblastine,oxorubicin, carmustine (BCNU), lomustine (CCNU), cytarabine USP,cyclophosphamide, estramucine phosphate sodium, altretamine,hydroxyurea, ifosfamide, procarbazine, mitomycin, busulfan,cyclophosphamide, mitoxantrone, carboplatin, cisplatin, interferonalfa-2a recombinant, paclitaxel, teniposide, and streptozoci), cytotoxicalkylating agents (e.g., busulfan, chlorambucil, cyclophosphamide,melphalan, or ethylesulfonic acid), alkylating agents (e.g., asaley,AZQ, BCNU, busulfan, bisulphan, carboxyphthalatoplatinum, CBDCA, CCNU,CHIP, chlorambucil, chlorozotocin, cis-platinum, clomesone,cyanomorpholinodoxorubicin, cyclodisone, cyclophosphamide,dianhydrogalactitol, fluorodopan, hepsulfam, hycanthone, iphosphamide,melphalan, methyl CCNU, mitomycin C, mitozolamide, nitrogen mustard,PCNU, piperazine, piperazinedione, pipobroman, porfiromycin,spirohydantoin mustard, streptozotocin, teroxirone, tetraplatin,thiotepa, triethylenemelamine, uracil nitrogen mustard, and Yoshi-864),antimitotic agents (e.g., allocolchicine, Halichondrin M, colchicine,colchicine derivatives, dolastatin 10, maytansine, rhizoxin, paclitaxelderivatives, paclitaxel, thiocolchicine, trityl cysteine, vinblastinesulfate, and vincristine sulfate), plant alkaloids (e.g., actinomycin D,bleomycin, L-asparaginase, idarubicin, vinblastine sulfate, vincristinesulfate, mitramycin, mitomycin, daunorubicin, VP-16-213, VM-26,navelbine and taxotere), biologicals (e.g., alpha interferon, BCG,G-CSF, GM-CSF, and interleukin-2), topoisomerase I inhibitors (e.g.,camptothecin, camptothecin derivatives, and morpholinodoxorubicin),topoisomerase II inhibitors (e.g., mitoxantron, amonafide, m-AMSA,anthrapyrazole derivatives, pyrazoloacridine, bisantrene HCL,daunorubicin, deoxydoxorubicin, menogaril, N,N-dibenzyl daunomycin,oxanthrazole, rubidazone, VM-26 and VP-16), and synthetics (e.g.,hydroxyurea, procarbazine, o,p′-DDD, dacarbazine, CCNU, BCNU,cis-diamminedichloroplatimun, mitoxantrone, CBDCA, levamisole,hexamethylmelamine, all-trans retinoic acid, gliadel and porfimersodium). In some embodiments, a composition of the invention isadministered before, during, or after administration of at least oneantiproliferative agent for the treatment of cancer. Antiproliferativeagents are compounds that decrease the proliferation of cells.Antiproliferative agents include, but are not limited to, alkylatingagents, antimetabolites, enzymes, biological response modifiers,miscellaneous agents, hormones and antagonists, androgen inhibitors(e.g., flutamide and leuprolide acetate), antiestrogens (e.g., tamoxifencitrate and analogs thereof, toremifene, droloxifene and roloxifene),Additional examples of specific antiproliferative agents include, butare not limited to levamisole, gallium nitrate, granisetron,sargramostim strontium-89 chloride, filgrastim, pilocarpine,dexrazoxane, and ondansetron.

The compositions of the invention can be administered alone or incombination with other anti-tumor agents, includingcytotoxic/antineoplastic agents and anti-angiogenic agents.Cytotoxic/anti-neoplastic agents are defined as agents which attack andkill cancer cells. Some cytotoxic/anti-neoplastic agents are alkylatingagents, which alkylate the genetic material in tumor cells, e.g.,cis-platin, cyclophosphamide, nitrogen mustard, trimethylenethiophosphoramide, carmustine, busulfan, chlorambucil, belustine, uracilmustard, chlomaphazin, and dacabazine. Other cytotoxic/anti-neoplasticagents are antimetabolites for tumor cells, e.g., cytosine arabinoside,fluorouracil, methotrexate, mercaptopuirine, azathioprime, andprocarbazine. Other cytotoxic/anti-neoplastic agents are antibiotics,e.g., doxorubicin, bleomycin, dactinomycin, daunorubicin, mithramycin,mitomycin, mytomycin C, and daunomycin. There are numerous liposomalformulations commercially available for these compounds. Still othercytotoxic/anti-neoplastic agents are mitotic inhibitors (vincaalkaloids). These include vincristine, vinblastine and etoposide.Miscellaneous cytotoxic/anti-neoplastic agents include taxol and itsderivatives, L-asparaginase, anti-tumor antibodies, dacarbazine,azacytidine, amsacrine, melphalan, VM-26, ifosfamide, mitoxantrone, andvindesine.

Anti-angiogenic agents are well known to those of skill in the art.Suitable anti-angiogenic agents for combining with the compositions ofthe present disclosure include anti-VEGF antibodies, including humanizedand chimeric antibodies, anti-VEGF aptamers and antisenseoligonucleotides. Other known inhibitors of angiogenesis includeangiostatin, endostatin, interferons, interleukin 1 (including alpha andbeta) interleukin 12, retinoic acid, and tissue inhibitors ofmetalloproteinase-1 and -2. (TIMP-1 and -2). Small molecules, includingtopoisomerases such as razoxane, a topoisomerase II inhibitor withanti-angiogenic activity, can also be used.

Other anti-cancer agents that can be used in combination with thecompositions of the invention include, but are not limited to: acivicin;aclarubicin; acodazole hydrochloride; acronine; adozelesin; aldesleukin;altretamine; ambomycin; ametantrone acetate; aminoglutethimide;amsacrine; anastrozole; anthramycin; asparaginase; asperlin;azacitidine; azetepa; azotomycin; batimastat; benzodepa; bicalutamide;bisantrene hydrochloride; bisnafide dimesylate; bizelesin; bleomycinsulfate; brequinar sodium; bropirimine; busulfan; cactinomycin;calusterone; caracemide; carbetimer; carboplatin; carmustine; carubicinhydrochloride; carzelesin; cedefingol; chlorambucil; cirolemycin;cisplatin; cladribine; crisnatol mesylate; cyclophosphamide; cytarabine;dacarbazine; dactinomycin; daunorubicin hydrochloride; decitabine;dexormaplatin; dezaguanine; dezaguanine mesylate; diaziquone; docetaxel;doxorubicin; doxorubicin hydrochloride; droloxifene; droloxifenecitrate; dromostanolone propionate; duazomycin; edatrexate; eflornithinehydrochloride; elsamitrucin; enloplatin; enpromate; epipropidine;epirubicin hydrochloride; erbulozole; esorubicin hydrochloride;estramustine; estramustine phosphate sodium; etanidazole; etoposide;etoposide phosphate; etoprine; fadrozole hydrochloride; fazarabine;fenretinide; floxuridine; fludarabine phosphate; fluorouracil;fluorocitabine; fosquidone; fostriecin sodium; gemcitabine; gemcitabinehydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide;ilmofosine; interleukin II (including recombinant interleukin II, orrIL2), interferon alfa-2a; interferon alfa-2b; interferon alfa-n1;interferon alfa-n3; interferon beta-I a; interferon gamma-I b;iproplatin; irinotecan hydrochloride; lanreotide acetate; letrozole;leuprolide acetate; liarozole hydrochloride; lometrexol sodium;lomustine; losoxantrone hydrochloride; masoprocol; maytansine;mechlorethamine hydrochloride; megestrol acetate; melengestrol acetate;melphalan; menogaril; mercaptopurine; methotrexate; methotrexate sodium;metoprine; meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin;mitomalcin; mitomycin; mitosper; mitotane; mitoxantrone hydrochloride;mycophenolic acid; nocodazole; nogalamycin; ormaplatin; oxisuran;paclitaxel; pegaspargase; peliomycin; pentamustine; peplomycin sulfate;perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride;plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine;procarbazine hydrochloride; puromycin; puromycin hydrochloride;pyrazofurin; riboprine; rogletimide; safingol; safingol hydrochloride;semustine; simtrazene; sparfosate sodium; sparsomycin; spirogermaniumhydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin;sulofenur; talisomycin; tecogalan sodium; tegafur; teloxantronehydrochloride; temoporfin; teniposide; teroxirone; testolactone;thiamiprine; thioguanine; thiotepa; tiazofurin; tirapazamine; toremifenecitrate; trestolone acetate; triciribine phosphate; trimetrexate;trimetrexate glucuronate; triptorelin; tubulozole hydrochloride; uracilmustard; uredepa; vapreotide; verteporfin; vinblastine sulfate;vincristine sulfate; vindesine; vindesine sulfate; vinepidine sulfate;vinglycinate sulfate; vinleurosine sulfate; vinorelbine tartrate;vinrosidine sulfate; vinzolidine sulfate; vorozole; zeniplatin;zinostatin; zorubicin hydrochloride. Other anti-cancer drugs include,but are not limited to: 20-epi-1,25 dihydroxyvitamin D3;5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol;adozelesin; aldesleukin; ALL-TK antagonists; altretamine; ambamustine;amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine;anagrelide; anastrozole; andrographolide; angiogenesis inhibitors;antagonist D; antagonist G; antarelix; anti-dorsalizing morphogeneticprotein-1; antiandrogen, prostatic carcinoma; antiestrogen;antineoplaston; antisense oligonucleotides; aphidicolin glycinate;apoptosis gene modulators; apoptosis regulators; apurinic acid;ara-CDP-DL-PTBA; arginine deaminase; asulacrine; atamestane;atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron;azatoxin; azatyrosine; baccatin III derivatives; balanol; batimastat;BCR/ABL antagonists; benzochlorins; benzoylstaurosporine; beta lactamderivatives; beta-alethine; betaclamycin B; betulinic acid; bFGFinhibitor; bicalutamide; bisantrene; bisaziridinylspermine; bisnafide;bistratene A; bizelesin; breflate; bropirimine; budotitane; buthioninesulfoximine; calcipotriol; calphostin C; camptothecin derivatives;canarypox IL-2; capecitabine; carboxamide-amino-triazole;carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor;carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropinB; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost;cis-porphyrin; cladribine; clomifene analogues; clotrimazole;collismycin A; collismycin B; combretastatin A4; combretastatinanalogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8;cryptophycin A derivatives; curacin A; cyclopentanthraquinones;cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor;cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin;dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone;didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine;dihydrotaxol, 9-; dioxamycin; diphenyl spiromustine; docetaxel;docosanol; dolasetron; doxifluridine; droloxifene; dronabinol;duocarmycin SA; ebselen; ecomustine; edelfosine; edrecolomab;eflornithine; elemene; emitefur; epirubicin; epristeride; estramustineanalogue; estrogen agonists; estrogen antagonists; etanidazole;etoposide phosphate; exemestane; fadrozole; fazarabine; fenretinide;filgrastim; finasteride; flavopiridol; flezelastine; fluasterone;fludarabine; fluorodaunorunicin hydrochloride; forfenimex; formestane;fostriecin; fotemustine; gadolinium texaphyrin; gallium nitrate;galocitabine; ganirelix; gelatinase inhibitors; gemcitabine; glutathioneinhibitors; hepsulfam; heregulin; hexamethylene bisacetamide; hypericin;ibandronic acid; idarubicin; idoxifene; idramantone; ilmofosine;ilomastat; imidazoacridones; imiquimod; immunostimulant peptides;insulin-like growth factor-1 receptor inhibitor; interferon agonists;interferons; interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-;iroplact; irsogladine; isobengazole; isohomohalicondrin B; itasetron;jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide;leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole;leukemia inhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine;lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides;maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysininhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone;meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone;miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone;mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growthfactor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonalantibody, human chorionic gonadotrophin; monophosphoryl lipidA+myobacterium cell wall sk; mopidamol; multiple drug resistance geneinhibitor; multiple tumor suppressor 1-based therapy; mustard anticanceragent; mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; neutral endopeptidase; nilutamide;nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn;O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; paclitaxel; paclitaxel analogues;paclitaxel derivatives; palauamine; palmitoylrhizoxin; pamidronic acid;panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase;peldesine; pentosan polysulfate sodium; pentostatin; pentrozole;perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenylacetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum compounds;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RII retinamide; rogletimide;rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl; safingol;saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics;semustine; senescence derived inhibitor 1; sense oligonucleotides;signal transduction inhibitors; signal transduction modulators; singlechain antigen binding protein; sizofuran; sobuzoxane; sodiumborocaptate; sodium phenylacetate; solverol; somatomedin bindingprotein; sonermin; sparfosic acid; spicamycin D; spiromustine;splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-celldivision inhibitors; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; synthetic glycosaminoglycans; tallimustine;tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium;tegafur; tellurapyrylium; telomerase inhibitors; temoporfin;temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; totipotent stem cell factor;translation inhibitors; tretinoin; triacetyluridine; triciribine;trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinaseinhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenitalsinus-derived growth inhibitory factor; urokinase receptor antagonists;vapreotide; variolin B; vector system, erythrocyte gene therapy;velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine;vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; and zinostatinstimalamer. In one embodiment, the anti-cancer drug is 5-fluorouracil,taxol, or leucovorin.

Anti-Viral Agents

The present invention contemplates compositions comprising at least oneSCFA or compound comprising a SCFA moiety as described herein incombination with anti-viral agents. Anti-viral agents include, but arenot limited to, inhibitors of viral uncoating (e.g., amantadine andrimantidine), reverse transcriptase inhibitors (e.g., acyclovir,zidovudine, and lamivudine), agents that target integrase; agents thatblock attachment of transcription factors to viral DNA; agents (e.g.,antisense molecules) that impact translation (e.g., fomivirsen); agentsthat modulate translation/ribozyme function; protease inhibitors; viralassembly modulators (e.g., rifampicin); antiretrovirals such as, forexample, nucleoside analogue reverse transcriptase inhibitors (e.g.,azidothymidine (AZT), ddl, ddC, 3TC, d4T); non-nucleoside reversetranscriptase inhibitors (e.g., efavirenz, nevirapine); nucleotideanalogue reverse transcriptase inhibitors; and agents that preventrelease of viral particles (e.g., zanamivir and oseltamivir), abacavir,adefovir, amantadine, amprenavir, ampligen, arbidol, atazanavir,atripla, boceprevirertet, cidofovir, combivir, darunavir, delavirdine,didanosine, docosanol, edoxudine, emtricitabine, enfuvirtide, entecavir,famciclovir, fosamprenavir, foscarnet, fosfonet, ganciclovir,ibacitabine, imunovir, idoxuridine, imiquimod, indinavir, inosine,various interferons (e.g., peginterferon alfa-2a), lopinavir, loviride,maraviroc, moroxydine, methisazone, nelfinavir, nexavir, penciclovir,peramivir, pleconaril, podophyllotoxin, raltegravir, ribavirin,ritonavir, pyramidine, saquinavir, stavudine, telaprevir, tenofovir,tipranavir, trifluridine, trizivir, tromantadine, truvada, valaciclovir,valganciclovir, vicriviroc, vidarabine, viramidine, and zalcitabine.

Antiparasitic Agents

The present invention contemplates compositions comprising at least oneSCFA or compound comprising a SCFA moiety as described herein incombination with antiparasitic agents. Such agents include, but are notlimited to, thiabendazole, pyrantel pamoate, mebendazole, praziquantel,niclosamide, bithionol, oxamniquine, metrifonate, ivermectin,albendazole, eflornithine, melarsoprol, pentamidine, benznidazole,nifurtimox, and nitroimidazole. The skilled artisan is aware of otheragents that may find utility for the treatment of parasitic disorders.

Antibacterial Agents

The present invention contemplates compositions comprising at least oneSCFA or compound comprising a SCFA moiety as described herein incombination with agents useful in the treatment or prevention ofbacterial disorders. Antibacterial agents can be classified in variousmanners, including based on mechanism of action, based on chemicalstructure, and based on spectrum of activity. Examples of antibacterialagents include those that target the bacterial cell wall (e.g.,cephalosporins and penicillins) or the cell membrane (e.g., polymyxins),or interfere with essential bacterial enzymes (e.g., sulfonamides,rifamycins, and quinolines). Most antibacterial agents that targetprotein synthesis (e.g., tetracyclines and macrolides) arebacteriostatic, whereas agents such as the aminoglycoside arebactericidal. Another means of categorizing antibacterial agents isbased on their target specificity; “narrow-spectrum” agents targetspecific types of bacteria (e.g., Gram-positive bacteria such asStreptococcus), while “broad-spectrum” agents have activity against abroader range of bacteria. The skilled artisan is aware of types ofanti-bacterial agents that are appropriate for use in specific bacterialinfections.

Methods of Treatment

One aspect of the invention relates to a treatment regimen for treatingor preventing a disease or disorder using a composition of theinvention. Administration of the compositions of the present inventionto a subject may be carried out using known procedures, at dosages andfor periods of time effective for treating or preventing a disease ordisorder in the subject. An effective amount of the therapeutic compoundnecessary to achieve a therapeutic effect may vary according to factorssuch as the state of the disease or disorder in the subject; the age,sex, and weight of the subject.

The regimen of administration may affect what constitutes an effectiveamount. Further, the dosages of the compositions may be proportionallyincreased or decreased as indicated by the exigencies of the therapeuticor prophylactic situation. A non-limiting example of an effective doserange for a therapeutic compound of the invention is from about 1 toabout 5,000 mg/kg of body weight/per day. The effective dose can bebetween 1 μg to 10 mg active component/kg body weight/time, and can be20 μg to 10 mg component/kg body weight/time. The therapeutic compoundcan be administered every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31days. The number of doses for effective treatment can be 1, 2, 3, 4, 5,6, 7, 8, 9, 10 or more than 10.

One of ordinary skill in the art would be able to study the relevantfactors and make the determination regarding the effective amount of thetherapeutic compound without undue experimentation.

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions of this invention may be varied so as to obtain an amountof the active ingredient that is effective to achieve the desiredtherapeutic response for a particular subject, composition, and mode ofadministration, without being toxic to the subject.

In particular, the selected dosage level will depend upon a variety offactors including the activity of the particular compound employed, thetime of administration, the rate of excretion of the compound, theduration of the treatment, other drugs, compounds or materials used incombination with the compound, the age, sex, weight, condition, generalhealth and prior medical history of the subject being treated, and likefactors well known in the medical arts.

A medical doctor, e.g., physician or veterinarian, having ordinary skillin the art may readily determine and prescribe the effective amount ofthe pharmaceutical composition required. For example, the physician orveterinarian could start doses of the compounds of the inventionemployed in the pharmaceutical composition at levels lower than thatrequired in order to achieve the desired therapeutic effect andgradually increase the dosage until the desired effect is achieved.

Compositions of the invention for administration may be in the range offrom about 1 μg to about 10,000 mg, about 20 μg to about 9,500 mg, about40 μg to about 9,000 mg, about 75 μg to about 8,500 mg, about 150 μg toabout 7,500 mg, about 200 μg to about 7,000 mg, about 3050 μg to about6,000 mg, about 500 μg to about 5,000 mg, about 750 μg to about 4,000mg, about 1 mg to about 3,000 mg, about 10 mg to about 2,500 mg, about20 mg to about 2,000 mg, about 25 mg to about 1,500 mg, about 50 mg toabout 1,000 mg, about 75 mg to about 900 mg, about 100 mg to about 800mg, about 250 mg to about 750 mg, about 300 mg to about 600 mg, about400 mg to about 500 mg, and any and all whole or partial incrementstherebetween.

In some embodiments, the dose of a compound of the invention is fromabout 1 mg and about 2,500 mg. In some embodiments, a dose of a compoundof the invention used in compositions described herein is less thanabout 10,000 mg, or less than about 8,000 mg, or less than about 6,000mg, or less than about 5,000 mg, or less than about 3,000 mg, or lessthan about 2,000 mg, or less than about 1,000 mg, or less than about 500mg, or less than about 200 mg, or less than about 50 mg, or less thanabout 40 mg, or less than about 30 mg, or less than about 25 mg, or lessthan about 20 mg, or less than about 15 mg, or less than about 10 mg, orless than about 5 mg, or less than about 2 mg, or less than about 1 mg,or less than about 0.5 mg, and any and all whole or partial incrementsthere between.

In one embodiment, the treatment regimen comprises daily administrationof a composition of the invention. In one embodiment, a treatmentregimen comprises administering a short chain fatty acid at least oncedaily for at least 1 day, at least 2 days, at least 3 days, at least 4days, at least 5 days, at least 7 days, at least 10 days, at least 2weeks, at least 3 weeks, at least 1 month, at least 2 months, at least 3months, at least 6 months, at least 1 year or more than 1 year. In oneembodiment, a treatment regimen comprises administering a short chainfatty acid three times daily for at least 1 day, at least 2 days, atleast 3 days, at least 4 days, at least 5 days, at least 7 days, atleast 10 days, at least 2 weeks, at least 3 weeks, at least 1 month, atleast 2 months, at least 3 months, at least 6 months, at least 1 year ormore than 1 year. In one embodiment, a treatment regimen comprisesadministering a composition comprising a short chain fatty acid, a shortchain fatty acid precursor, a short chain fatty acid biosynthesisprecursor, or a combination thereof at least once daily upon appearanceof a disease or disorder.

In one embodiment, a treatment regimen comprises daily oraladministration of a short chain fatty acid. In one exemplary embodiment,600 mg of butyrate is administered 3 times per day (a total of 1800mg/day) for at least one week. In one embodiment, a treatment regimencomprises oral administration of two butyrate capsules containing 600 mgof butyrate 3 times per day (a total of 3600 mg/day) for at least oneweek. In one embodiment, a treatment regimen comprises oraladministration of two butyrate capsules containing 600 mg of butyrate 3times per day (a total of 3600 mg/day) for at least one week followedthereafter by oral administration of butyrate capsules containing 600 mgof butyrate 3 times per day (a total of 1800 mg/day) for at least oneweek.

As described herein, administration of a composition of the invention toan individual makes it possible to induce tolerance, strengthen gutbarrier integrity, and reduce inflammation in the individual. The methodcomprises administering to an individual a composition described herein.In some embodiments, the composition comprises one or more SCFAs or SCFAderivatives. The composition is administered to the individual insufficient quantity to produce the desired effect of inducing tolerance,strengthening the gut barrier, and reducing inflammation. It may beadministered to an individual in need of treatment, reduction in theseverity of or prevention of at least one disease selected from anautoimmune disease, an inflammatory disease, an allergic disease, or aninfectious disease.

Whether administration of the composition induces tolerance can bedetermined by using, as an index, increase or reinforcement of at leastone of the following: the number of regulatory T cells (Tregs), theratio of Tregs in the T cell group of the colon, a function of Tregs, orexpression of a marker of Tregs. A specific approach is measurementcounts or percentage of Foxp3-expressing Tregs in a patient sample, suchas a biopsy or a blood sample, promotion (enhancement) of IL-10expression, promotion (enhancement) of CTLA4 expression, promotion(enhancement) of IDO expression, or suppression of IL-4 expression asthe index of the induction of proliferation or accumulation ofregulatory T cells. Whether administration of the compositionstrengthens barrier function can be determined by using, as an index,increase in production of active form TGF-β and/or tightjunction-related proteins by intestinal epithelial cells. Whetheradministration of the composition reduces inflammation can be determinedby using, as an index, increase in production of anti-inflammatorycytokines such as IL-10 and/or TGF-.beta., or decrease in production ofpro-inflammatory cytokines such as IL-4.

Methods for detecting such expression include northern blotting, RT-PCR,and dot blotting for detection of gene expression at the transcriptionlevel; ELISA, radioimmunoassays, immunoblotting, immunoprecipitation,and flow cytometry for detection of gene expression at the translationlevel. Samples that may be used for measuring such an index includetissues and fluids obtained from an individual, such as blood, a biopsy,or a fecal sample.

Pharmaceutical Compositions

The present invention includes pharmaceutical compositions comprisingone or more compositions of the present invention. The formulations ofthe pharmaceutical compositions described herein may be prepared by anymethod known or hereafter developed in the art of pharmacology. Ingeneral, such preparatory methods include the step of bringing theactive ingredient into association with a carrier or one or more otheraccessory ingredients, and then, if necessary or desirable, shaping orpackaging the product into a desired single- or multi-dose unit.

Said compositions may comprise additional medicinal agents,pharmaceutical agents, carriers, buffers, adjuvants, dispersing agents,diluents, and the like depending on the intended use and application.

Examples of suitable pharmaceutical carriers, excipients and/or diluentsare well known in the art and include, but are not limited to, a gum, astarch (e g. corn starch, pre-gelatinized starch), a sugar (e.g.,lactose, mannitol, sucrose, dextrose), a cellulosic material (e.g.microcrystalline cellulose), an acrylate (e.g. polymethylacrylate),calcium carbonate, magnesium oxide, talc, or mixtures thereof.

Pharmaceutically acceptable carriers for liquid formulations are aqueousor non-aqueous solutions, suspensions, emulsions or oils, Examples ofnon-aqueous solvents are propylene glycol, polyethylene glycol, andinjectable organic esters such as ethyl oleate. Examples of oils arethose of animal, vegetable, or synthetic origin, for example, peanutoil, soybean oil, olive oil, sunflower oil, turmeric oil, fish-liveroil, another marine oil, or a lipid from milk or eggs.

Aqueous carriers include water, alcoholic/aqueous solutions, emulsionsor suspensions, including saline and buffered media such as phosphatebuffered saline solutions, water, emulsions, such as oil/wateremulsions, various types of wetting agents, sterile solutions etc.Compositions comprising such carriers can be formulated by well-knownconventional methods. Suitable carriers may comprise any material which,when combined with the biologically active compound of the invention,retains the biological activity. Preparations for parenteraladministration may include sterile aqueous or non-aqueous solutions,suspensions, and emulsions. Examples of non-aqueous solvents arepropylene glycol, polyethylene glycol, vegetable oils such as olive oil,and injectable organic esters such as ethyl oleate. Aqueous carriersinclude water, alcoholic/aqueous solutions, emulsions or suspensions,including saline and buffered media. Parenteral vehicles may includesodium chloride solution, Ringer's dextrose, dextrose and sodiumchloride, lactated Ringer's, or fixed oils. Intravenous vehicles mayinclude fluid and nutrient replenishes, electrolyte replenishers (suchas those based on Ringer's dextrose), and the like. Preservatives andother additives may also be present including, for example,antimicrobials, anti-oxidants, chelating agents, and inert gases and thelike, in addition, the pharmaceutical composition of the presentinvention might comprise proteinaceous carriers, like, e.g., serumalbumin or immunoglobulin.

The pharmaceutical compositions provided herein may also be administeredas controlled-release compositions, i.e. compositions in which theactive ingredient is released over a period of time afteradministration. Controlled- or sustained-release compositions includeformulation in lipophilic depots (e.g. fatty acids, waxes, oils). Inanother embodiment, the composition is an immediate-release composition,i.e. a composition in which all the active ingredient is releasedimmediately after administration.

Further, the pharmaceutical compositions according to the invention andas described herein in the various embodiments may or a compositioncomprising said compound may be administered admixed to food, functionalfood, drinks, medicinal food.

Although the description of pharmaceutical compositions provided hereinare principally directed to pharmaceutical compositions which aresuitable for ethical administration to humans, it will be understood bythe skilled artisan that such compositions are generally suitable foradministration to animals of all sorts. Modification of pharmaceuticalcompositions suitable for administration to humans in order to renderthe compositions suitable for administration to various animals is wellunderstood, and the ordinarily skilled veterinary pharmacologist candesign and perform such modification with merely ordinary, if any,experimentation. Subjects to which administration of the pharmaceuticalcompositions of the invention are contemplated include, but are notlimited to, humans and other primates, mammals including commerciallyrelevant mammals such as non-human primates, cattle, pigs, horses,sheep, cats, and dogs, non-mammalian animals, and other veterinaryapplications.

Pharmaceutical compositions that are useful in the methods of theinvention may be prepared, packaged, or sold in formulations suitablefor ophthalmic, oral, rectal, vaginal, parenteral, topical, pulmonary,intranasal, buccal, intratumoral, epidural, intracerebral,intracerebroventricular, or another route of administration. Othercontemplated formulations include projected nanoparticles, liposomalpreparations, resealed erythrocytes containing the active ingredient,and immunologically-based formulations.

A pharmaceutical composition of the invention may be prepared, packaged,or sold in bulk, as a single unit dose, or as a plurality of single unitdoses. As used herein, a “unit dose” is discrete amount of thepharmaceutical composition comprising a predetermined amount of theactive ingredient. The amount of the active ingredient is generallyequal to the dosage of the active ingredient which would be administeredto a subject or a convenient fraction of such a dosage such as, forexample, one-half or one-third of such a dosage.

The relative amounts of the active ingredient, the pharmaceuticallyacceptable carrier, and any additional ingredients in a pharmaceuticalcomposition of the invention will vary, depending upon the identity,size, and condition of the subject treated and further depending uponthe route by which the composition is to be administered. By way ofexample, the composition may comprise between 0.1% and 100% (w/w) activeingredient.

In addition to the active ingredient, a pharmaceutical composition ofthe invention may further comprise one or more additionalpharmaceutically active agents.

Controlled- or sustained-release formulations of a pharmaceuticalcomposition of the invention may be made using conventional technology.

Pharmaceutical compositions also include nutritional compositions, suchas oral nutritional compositions for oral consumption and optionally forenteral adsorption, wherein the nutritional composition includes thecompounds of the present invention. Therefore, in one embodiment, theinvention relates to nutraceutical compositions.

If the nutritional compositions are formulated to be administeredorally, the compositions may be a liquid oral nutritional supplement(e.g., incomplete feeding) or a complete feeding. In this manner, thenutritional compositions may be administered in any known formincluding, for example, tablets, capsules, liquids, chewables, softgels, sachets, powders, syrups, liquid suspensions, emulsions, infantformulas and solutions in convenient dosage forms.

A nutritional formula encompasses any nutritionally complete orsupplementary formulation (a nutritional supplement, for example). Asused herein, “nutritionally complete” are preferably nutritionalproducts that contain sufficient types and levels of macronutrients(protein, fats and carbohydrates) and micronutrients to be sufficient tobe a sole source of nutrition for the subject to which it is beingadministered to. Patients can receive 100% of their nutritionalrequirements from such complete nutritional compositions. According toone embodiment, the nutritional formula is a supplementary formulationproviding supplementary nutrition. A “supplementary formula” may not benutritionally complete, but preferably contains specific nutrients thatare supportive, for example in combination with physical exercise, withfurther of the beneficial effects of the invention, and/or which addressspecific or additional needs of the subject.

The nutritional formula may be a generally applicable nutritionalformula, for example adapted to subjects of a specific age, for examplea formula for children, but it may also be a formula for elderlypatients, for intensive care patients, or a specially adapted formulafor patients suffering from a specific disease, for example. Anynutritional formula may be reconstitutable, that is, present in asubstantially dried, for example powdered form, or ready-to-drink, inthe form of liquid formulas, for example.

Formulations of a pharmaceutical composition suitable for parenteraladministration comprise the active ingredient combined with apharmaceutically acceptable carrier, such as sterile water or sterileisotonic saline. Such formulations may be prepared, packaged, or sold ina form suitable for bolus administration or for continuousadministration. Injectable formulations may be prepared, packaged, orsold in unit dosage form, such as in ampules or in multi dose containerscontaining a preservative. Formulations for parenteral administrationinclude, but are not limited to, suspensions, solutions, emulsions inoily or aqueous vehicles, pastes, and implantable sustained-release orbiodegradable formulations. Such formulations may further comprise oneor more additional ingredients including, but not limited to,suspending, stabilizing, or dispersing agents. In one embodiment of aformulation for parenteral administration, the active ingredient isprovided in dry (i.e., powder or granular) form for reconstitution witha suitable vehicle (e.g., sterile pyrogen free water) prior toparenteral administration of the reconstituted composition.

The pharmaceutical compositions may be prepared, packaged, or sold inthe form of a sterile injectable aqueous or oily suspension or solution.This suspension or solution may be formulated according to the knownart, and may comprise, in addition to the active ingredient, additionalingredients such as the dispersing agents, wetting agents, or suspendingagents described herein. Such sterile injectable formulations may beprepared using a non-toxic parenterally acceptable diluent or solvent,such as water or 1,3 butane diol, for example. Other acceptable diluentsand solvents include, but are not limited to, Ringer's solution,isotonic sodium chloride solution, and fixed oils such as synthetic monoor di-glycerides. Other parentally-administrable formulations which areuseful include those which comprise the active ingredient inmicrocrystalline form, in a liposomal preparation, or as a component ofa biodegradable polymer system. Compositions for sustained release orimplantation may comprise pharmaceutically acceptable polymeric orhydrophobic materials such as an emulsion, an ion exchange resin, asparingly soluble polymer, or a sparingly soluble salt.

The compositions of the present invention as described herein may beused as a complete food product, as a component of a food product, as adietary supplement or as part of a dietary supplement, as a feedadditive and may be either in liquid, semisolid or solid form. Thecompositions of the present invention as described herein additionallymay be in the form of a pharmaceutical composition. The compositions,dietary supplements, food products, baby food products, feed additives,and/or pharmaceutical compositions of the present invention mayadvantageously be utilized in methods for promoting the health of anindividual.

As indicated above, the compositions may be in liquid, semisolid orsolid form. For example, the compositions may be administered astablets, gel packs, capsules, gelatin capsules, flavored drinks, as apowder that can be reconstituted into such a drink, cooking oil, saladoil or dressing, sauce, syrup, mayonnaise, margarine or the like.Furthermore, the food product, dietary supplements, and the like, of thepresent invention can include, but are not limited to, dairy products,baby food, baby formula, beverages, bars, a powder, a food topping, adrink, a cereal, an ice cream, a candy, a snack mix, a baked foodproduct and a fried food product. Beverages of the invention include butare not limited to energy drinks, nutraceutical drinks, smoothies,sports drinks, orange juice and other fruit drinks. A bar of the presentinvention includes, but is not limited to, a meal replacement, anutritional bar, a snack bar and an energy bar, an extruded bar, and thelike. Dairy products of the invention include, but are not limited to,including but not limited to yogurt, yogurt drinks, cheese and milk.

The food products or dietary supplements of the present invention mayfurther comprise herbals, herbal extracts, fungal extracts, enzymes,fiber sources, minerals, and vitamins. The microalgal oils andmicroalgal biomass of the present invention may be used in thecompositions of the invention for both therapeutic and non-therapeuticuses. Thus, the compositions, food products and animal feed additives ofthe present invention may be used for therapeutic or non-therapeuticpurposes.

Compositions intended for oral administration may be prepared accordingto any known method for the manufacture of dietary supplements orpharmaceutical preparations, and such compositions may include at leastone additive selected from the group consisting of taste improvingsubstances, such as sweetening agents or flavoring agents, stabilizers,emulsifiers, coloring agents and preserving agents in order to provide adietetically or pharmaceutically palatable preparation. Vitamins,minerals and trace element from any physiologically acceptable sourcemay also be included in the composition of the invention.

A pharmaceutical composition of the present invention comprises the saidcompositions of the present invention in a therapeutically effectiveamount. The compositions may additionally comprise prescriptionmedicines or non-prescription medicines. The combinations mayadvantageously produce one or more of the following effects: (1)additive and/or synergistic benefits; (2) reduction of the side effectsand/or adverse effects associated with use of the prescription medicinein the absence of the said formulations; and/or (3) the ability to lowerthe dosage of the prescription medicine in comparison to the amount ofprescription medicine needed in the absence of the said formulations.

The active agents of the present invention can be prepared in the formof their pharmaceutically acceptable salts. As understood by one ofskill in the art, pharmaceutically acceptable salts are salts thatretain the desired biological activity of the parent compound and do notimpart undesired toxicological effects. “Pharmaceutically acceptablesalts” as defined herein include derivatives of the disclosed SCFA orcompound comprising a SCFA moiety, wherein the parent compound ismodified by making non-toxic salts of the carboxylate group thereof, andfurther refers to pharmaceutically acceptable hydrates, and solvates ofsuch compounds. Examples of pharmaceutically acceptable salts include,but are not limited to, mineral or organic acid salts of the carboxylicacid group of the SCFA. For example, conventional non-toxic acid saltsinclude those derived from inorganic acids such as hydrochloric,hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; andthe salts prepared from organic acids such as acetic, propionic,succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic,pamoic, maleic, hydroxylmaleic, phenylacetic, glutamic, benzoic,salicylic, mesylic, esylic, besylic, sulfanilic, 2-acetoxybenzoic,fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic,isethionic, HOOC—(CH₂)_(n)—COOH where n is 0-4, and the like. Lists ofadditional suitable salts may be found, e.g., in Remington'sPharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa.,p. 1418 (1985). In some embodiments the SCFA can be present as an esterof the SCFA's carboxylic acid with a branched or unbranched alkylalcohol of one to 6 carbons. For example, the SCFA can be present as anethyl ester, propyl ester, butyl ester, isopropyl ester, t-butyl ester,pentyl ester, or hexyl ester.

The active agents can be formulated for administration in accordancewith known pharmacy techniques. See, e.g., Remington, The Science AndPractice of Pharmacy (9th Ed. 1995). In the manufacture of apharmaceutical composition according to the present invention, theactive agents (including the physiologically acceptable salts thereof)is typically admixed with, inter alia, an acceptable carrier. Thecarrier must, of course, be acceptable in the sense of being compatiblewith any other ingredients in the formulation and must not bedeleterious to the subject. The carrier can be a solid or a liquid, orboth, and can be formulated with the active agent as a unit-doseformulation, for example, a tablet, which can contain from 0.01% or 0.5%to 95% or 99%, or any value between 0.01% and 99%, by weight of theactive agent. One or more active agents can be incorporated in thecompositions of the invention, which can be prepared by any of thewell-known techniques of pharmacy, comprising admixing the components,optionally including one or more accessory ingredients. Moreover, thecarrier can be preservative free, as described herein above.

In some embodiments, the active agents comprise a lower limit rangingfrom about 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 1, 2,3, 4, 5, 6, 7, 8, 9, and 10% to an upper limit ranging from about 10,11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28,29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46,47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64,65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 and100% by weight of the composition. In some embodiments, the active agentincludes from about 0.05% to about greater than 99% by weight of thecomposition.

The pharmaceutical compositions according to embodiments of the presentinvention are generally formulated for oral or topical (i.e., skin,ocular and mucosal surfaces) administration, with the most suitableroute in any given case depending on the nature and severity of thecondition being treated and on the nature of the particular active agentwhich is being used.

Topical Formulations

The compositions of the present invention and the pharmaceuticalcompositions containing said compounds, may be administered topically,and thus be formulated in a form suitable for topical administration,i.e. as a pH balanced cream preparation. An obstacle for topicaladministration of pharmaceuticals is the stratum corneum layer of theepidermis. The stratum corneum is a highly resistant layer comprised ofprotein, cholesterol, sphingolipids, free fatty acids and various otherlipids, and includes cornified and living cells. One of the factors thatlimit the penetration rate (flux) of a compound through the stratumcorneum is the amount of the active substance that can be loaded orapplied onto the skin surface. The greater the amount of activesubstance which is applied per unit of area of the skin, the greater theconcentration gradient between the skin surface and the lower layers ofthe skin, and in turn the greater the diffusion force of the activesubstance through the skin. Therefore, a formulation containing agreater concentration of the active substance is more likely to resultin penetration of the active substance through the skin, and more of it,and at a more consistent rate, than a formulation having a lesserconcentration, all other things being equal.

Formulations suitable for topical administration include, but are notlimited to, liquid or semi liquid preparations such as liniments,lotions, oil in water or water in oil emulsions such as creams,ointments or pastes, and solutions or suspensions. Further, formulationssuitable for topical administration can be in the form of cremes andliquids including, for example, syrups, suspensions or emulsions,inhalants, sprays, mousses, oils, gels, solids and the like. Topicallyadministrable formulations may, for example, comprise from about 1% toabout 10% (w/w) active ingredient, although the concentration of theactive ingredient may be as high as the solubility limit of the activeingredient in the solvent. Formulations for topical administration mayfurther comprise one or more of the additional ingredients describedherein.

Enhancers of permeation may be used. These materials increase the rateof penetration of drugs across the skin. Typical enhancers in the artinclude ethanol, glycerol monolaurate, PGML (polyethylene glycolmonolaurate), dimethylsulfoxide, and the like. Other enhancers includeoleic acid, oleyl alcohol, ethoxydiglycol, laurocapram, alkanecarboxylicacids, dimethylsulfoxide, polar lipids, or N-methyl-2-pyrrolidone.

One acceptable vehicle for topical delivery of some of the compositionsof the invention may contain liposomes. The composition of the liposomesand their use are known in the art (for example, see U.S. Pat. No.6,323,219).

In alternative embodiments, the topically active pharmaceuticalcomposition may be optionally combined with other ingredients such asadjuvants, anti-oxidants, chelating agents, surfactants, foaming agents,wetting agents, emulsifying agents, viscosifiers, buffering agents,preservatives, and the like. In another embodiment, a permeation orpenetration enhancer is included in the composition and is effective inimproving the percutaneous penetration of the active ingredient into andthrough the stratum corneum with respect to a composition lacking thepermeation enhancer. Various permeation enhancers, including oleic acid,oleyl alcohol, ethoxydiglycol, laurocapram, alkanecarboxylic acids,dimethylsulfoxide, polar lipids, or N-methyl-2-pyrrolidone, are known tothose of skill in the art. In another aspect, the composition mayfurther comprise a hydrotropic agent, which functions to increasedisorder in the structure of the stratum corneum, and thus allowsincreased transport across the stratum corneum. Various hydrotropicagents, such as isopropyl alcohol, propylene glycol, or sodium xylenesulfonate, are known to those of skill in the art.

The topically active pharmaceutical composition should be applied in anamount effective to affect desired changes. As used herein “amounteffective” shall mean an amount sufficient to cover the region of skinsurface where a change is desired. In various embodiments, an activecompound may be present in the amount of from about 0.0001% to about 15%by weight volume of the composition, from about 0.0005% to about 5% ofthe composition, or from about 0.001% to about 1% of the composition.Such compounds may be synthetically-or naturally derived.

Oral Formulations

The compositions of the present invention and the pharmaceuticalcompositions containing said compounds, may be administered orally, andthus be formulated in a form suitable for oral administration, i.e. as asolid or a liquid preparation. Suitable solid oral formulations includetablets, capsules, pills, granules, pellets and the like. Suitableliquid oral formulations include solutions, suspensions, dispersions,emulsions, oils and the like. If formulated in form of a capsule, thecompositions of the present invention comprise, in addition to theactive compound and the inert carrier or diluent, a hard gelatingcapsule. In one embodiment, a formulation for oral administration is anenteric coated, time release capsule.

Formulations suitable for oral administration can be presented indiscrete units, such as capsules, cachets, lozenges, or tablets, eachcontaining a predetermined amount of the active compound; as a powder orgranules; as a solution or a suspension in an aqueous or non-aqueousliquid; or as an oil-in-water or water-in-oil emulsion. Suchformulations can be prepared by any suitable method of pharmacy, whichincludes bringing into association the active compound and a suitablecarrier (which can contain one or more accessory ingredients as notedabove). In general, the formulations of the invention are prepared byuniformly and intimately admixing the active compound with a liquid orfinely divided solid carrier, or both, and then, if necessary, shapingthe resulting mixture. For example, a tablet can be prepared bycompressing or molding a powder or granules containing the activecompound, optionally with one or more accessory ingredients. Compressedtablets can be prepared by compressing, in a suitable machine, thecompound in a free-flowing form, such as a powder or granules optionallymixed with a binder, lubricant, inert diluent, and/or surfaceactive/dispersing agent(s). Molded tablets can be made by molding, in asuitable machine, the powdered compound moistened with an inert liquidbinder.

Formulations for Other Routes of Administration

The compositions of the present invention and the pharmaceuticalcompositions containing said compounds may be further administeredintranasally, i.e. by inhalation and thus may be formulated in a formsuitable for intranasal administration, i.e. as an aerosol or a liquidpreparation.

The compositions of the present invention may also, for example, beformulated for parenteral administration. Formulations of apharmaceutical composition suitable for parenteral administrationcomprise the active ingredient combined with a pharmaceuticallyacceptable carrier, such as sterile water or sterile isotonic saline.Such formulations may be prepared, packaged, or sold in a form suitablefor bolus administration or for continuous administration. Injectableformulations may be prepared, packaged, or sold in unit dosage form,such as in ampules or in multi dose containers containing apreservative. Formulations for parenteral administration include, butare not limited to, suspensions, solutions, emulsions in oily or aqueousvehicles, pastes, and implantable sustained-release or biodegradableformulations. Such formulations may further comprise one or moreadditional ingredients including, but not limited to, suspending,stabilizing, or dispersing agents. In one embodiment of a formulationfor parenteral administration, the active ingredient is provided in dry(i.e., powder or granular) form for reconstitution with a suitablevehicle (e.g., sterile pyrogen free water) prior to parenteraladministration of the reconstituted composition.

The pharmaceutical compositions may be prepared, packaged, or sold inthe form of a sterile injectable aqueous or oily suspension or solution.This suspension or solution may be formulated according to the knownart, and may comprise, in addition to the active ingredient, additionalingredients such as the dispersing agents, wetting agents, or suspendingagents described herein. Such sterile injectable formulations may beprepared using a non-toxic parenterally acceptable diluent or solvent,such as water or 1,3 butane diol, for example. Other acceptable diluentsand solvents include, but are not limited to, Ringer's solution,isotonic sodium chloride solution, and fixed oils such as synthetic monoor di-glycerides. Other parentally-administrable formulations which areuseful include those which comprise the active ingredient inmicrocrystalline form, in a liposomal preparation, or as a component ofa biodegradable polymer system. Compositions for sustained release orimplantation may comprise pharmaceutically acceptable polymeric orhydrophobic materials such as an emulsion, an ion exchange resin, asparingly soluble polymer, or a sparingly soluble salt.

Formulations are prepared using a pharmaceutically acceptable “carrier”composed of materials that are considered safe and effective and may beadministered to an individual without causing undesirable biologicalside effects or unwanted interactions. The “carrier” is all componentspresent in the pharmaceutical formulation other than the activeingredient or ingredients. The term “carrier” includes but is notlimited to diluents, binders, lubricants, desintegrators, fillers, andcoating compositions. “Carrier” also includes all components of thecoating composition which may include plasticizers, pigments, colorants,stabilizing agents, and glidants. The delayed release dosageformulations may be prepared as described in references such as“Pharmaceutical dosage form tablets”, eds. Liberman et. al. (New York,Marcel Dekker, Inc., 1989), “Remington—The science and practice ofpharmacy”, 20th ed., Lippincott Williams & Wilkins, Baltimore, Md.,2000, and “Pharmaceutical dosage forms and drug delivery systems”, 6thEdition, Ansel et. al., (Media, Pa.: Williams and Wilkins, 1995) whichprovides information on carriers, materials, equipment and process forpreparing tablets and capsules and delayed release dosage forms oftablets, capsules, and granules.

Examples of suitable coating materials include, but are not limited to,cellulose polymers such as cellulose acetate phthalate, hydroxypropylcellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulosephthalate and hydroxypropyl methylcellulose acetate succinate; polyvinylacetate phthalate, acrylic acid polymers and copolymers, and methacrylicresins that are commercially available under the trade name Eudragit®(Roth Pharma, Westerstadt, Germany), Zein, shellac, and polysaccharides.

Additionally, the coating material may contain conventional carrierssuch as plasticizers, pigments, colorants, glidants, stabilizationagents, pore formers and surfactants.

Optional pharmaceutically acceptable excipients present in thedrug-containing tablets, beads, granules or particles include, but arenot limited to, diluents, binders, lubricants, disintegrants, colorants,stabilizers, and surfactants. Diluents, also termed “fillers,” aretypically necessary to increase the bulk of a solid dosage form so thata practical size is provided for compression of tablets or formation ofbeads and granules. Suitable diluents include, but are not limited to,dicalcium phosphate dihydrate, calcium sulfate, lactose, sucrose,mannitol, sorbitol, cellulose, microcrystalline cellulose, kaolin,sodium chloride, dry starch, hydrolyzed starches, pregelatinized starch,silicone dioxide, titanium oxide, magnesium aluminum silicate and powdersugar.

Binders are used to impart cohesive qualities to a solid dosageformulation, and thus ensure that a tablet or bead or granule remainsintact after the formation of the dosage forms. Suitable bindermaterials include, but are not limited to, starch, pregelatinizedstarch, gelatin, sugars (including sucrose, glucose, dextrose, lactoseand sorbitol), polyethylene glycol, waxes, natural and synthetic gumssuch as acacia, tragacanth, sodium alginate, cellulose, includinghydroxypropylmethylcellulose, hydroxypropylcellulose, ethylcellulose,and veegum, and synthetic polymers such as acrylic acid and methacrylicacid copolymers, methacrylic acid copolymers, methyl methacrylatecopolymers, aminoalkyl methacrylate copolymers, polyacrylicacid/polymethacrylic acid and polyvinylpyrrolidone.

Lubricants are used to facilitate tablet manufacture. Examples ofsuitable lubricants include, but are not limited to, magnesium stearate,calcium stearate, stearic acid, glycerol behenate, polyethylene glycol,talc, and mineral oil.

Disintegrants are used to facilitate dosage form disintegration or“breakup” after administration, and generally include, but are notlimited to, starch, sodium starch glycolate, sodium carboxymethylstarch, sodium carboxymethylcellulose, hydroxypropyl cellulose,pregelatinized starch, clays, cellulose, alginine, gums or cross linkedpolymers, such as cross-linked PVP (Polyplasdone XL from GAF ChemicalCorp).

Stabilizers are used to inhibit or retard drug decomposition reactionswhich include, by way of example, oxidative reactions.

Surfactants may be anionic, cationic, amphoteric or nonionic surfaceactive agents. Suitable anionic surfactants include, but are not limitedto, those containing carboxylate, sulfonate and sulfate ions. Examplesof anionic surfactants include sodium, potassium, ammonium of long chainalkyl sulfonates and alkyl aryl sulfonates such as sodium dodecylbenzenesulfonate; dialkyl sodium sulfosuccinates, such as sodium dodecylbenzenesulfonate; dialkyl sodium sulfosuccinates, such as sodiumbis-(2-ethylthioxyl)-sulfosuccinate; and alkyl sulfates such as sodiumlauryl sulfate. Cationic surfactants include, but are not limited to,quaternary ammonium compounds such as benzalkonium chloride,benzethonium chloride, cetrimonium bromide, stearyl dimethylbenzylammonium chloride, polyoxyethylene and coconut amine. Examples ofnonionic surfactants include ethylene glycol monostearate, propyleneglycol myristate, glyceryl monostearate, glyceryl stearate,polyglyceryl-4-oleate, sorbitan acylate, sucrose acylate, PEG-150laurate, PEG-400 monolaurate, polyoxyethylene monolaurate, polysorbates,polyoxyethylene octylphenylether, PEG-1000 cetyl ether, polyoxyethylenetridecyl ether, polypropylene glycol butyl ether, Poloxamer® 401,stearoyl monoisopropanolamide, and polyoxyethylene hydrogenated tallowamide. Examples of amphoteric surfactants include sodiumN-dodecyl-beta-alanine, sodium N-lauryl-beta-iminodipropionate,myristoamphoacetate, lauryl betaine and lauryl sulfobetaine.

If desired, the tablets, beads granules or particles may also containminor amount of nontoxic auxiliary substances such as wetting oremulsifying agents, dyes, pH buffering agents, and preservatives.

As will be appreciated by those skilled in the art and as described inthe pertinent texts and literature, a number of methods are availablefor preparing drug-containing tablets, beads, granules or particles thatprovide a variety of drug release profiles. Such methods include, butare not limited to, the following: coating a drug or drug-containingcomposition with an appropriate coating material, typically although notnecessarily incorporating a polymeric material, increasing drug particlesize, placing the drug within a matrix, and forming complexes of thedrug with a suitable complexing agent.

The delayed release dosage units may be coated with the delayed releasepolymer coating using conventional techniques, e.g., using aconventional coating pan, an airless spray technique, fluidized bedcoating equipment (with or without a Wurster insert), or the like. Fordetailed information concerning materials, equipment and processes forpreparing tablets and delayed release dosage forms, see PharmaceuticalDosage Forms: Tablets, eds. Lieberman et al. (New York: Marcel Dekker,Inc., 1989), and Ansel et al., Pharmaceutical Dosage Forms and DrugDelivery Systems, 6^(th) Ed. (Media, Pa.: Williams & Wilkins, 1995).

An exemplary method for preparing extended release tablets is bycompressing a drug-containing blend, e.g., blend of granules, preparedusing a direct blend, wet-granulation, or dry-granulation process.Extended release tablets may also be molded rather than compressed,starting with a moist material containing a suitable water-solublelubricant. However, tablets are manufactured using compression ratherthan molding. A method for forming extended release drug-containingblend is to mix drug particles directly with one or more excipients suchas diluents (or fillers), binders, disintegrants, lubricants, glidants,and colorants. As an alternative to direct blending, a drug-containingblend may be prepared by using wet-granulation or dry-granulationprocesses. Beads containing the active agent may also be prepared by anyone of a number of conventional techniques, typically starting from afluid dispersion. For example, a typical method for preparingdrug-containing beads involves dispersing or dissolving the active agentin a coating suspension or solution containing pharmaceutical excipientssuch as polyvinylpyrrolidone, methylcellulose, talc, metallic stearates,silicone dioxide, plasticizers or the like. The admixture is used tocoat a bead core such as a sugar sphere (or so-called “non-pareil”)having a size of approximately 60 to 20 mesh.

An alternative procedure for preparing drug beads is by blending drugwith one or more pharmaceutically acceptable excipients, such asmicrocrystalline cellulose, lactose, cellulose, polyvinyl pyrrolidone,talc, magnesium stearate, a disintegrant, etc., extruding the blend,spheronizing the extrudate, drying and optionally coating to form theimmediate release beads.

Delayed release formulations are created by coating a solid dosage formwith a film of a polymer which is insoluble in the acid environment ofthe stomach, and soluble in the neutral environment of small intestines.The delayed release dosage units can be prepared, for example, bycoating a drug or a drug-containing composition with a selected coatingmaterial. The drug-containing composition may be, e.g., a tablet forincorporation into a capsule, a tablet for use as an inner core in a“coated core” dosage form, or a plurality of drug-containing beads,particles or granules, for incorporation into either a tablet orcapsule. Exemplary coating materials include bioerodible, graduallyhydrolyzable, gradually water-soluble, and/or enzymatically degradablepolymers, and may be conventional “enteric” polymers. Enteric polymers,as will be appreciated by those skilled in the art, become soluble inthe higher pH environment of the lower gastrointestinal tract or slowlyerode as the dosage form passes through the gastrointestinal tract,while enzymatically degradable polymers are degraded by bacterialenzymes present in the lower gastrointestinal tract, particularly in thecolon. Suitable coating materials for effecting delayed release include,but are not limited to, cellulosic polymers such as hydroxypropylcellulose, hydroxyethyl cellulose, hydroxymethyl cellulose,hydroxypropyl methyl cellulose, hydroxypropyl methyl cellulose acetatesuccinate, hydroxypropylmethyl cellulose phthalate, methylcellulose,ethyl cellulose, cellulose acetate, cellulose acetate phthalate,cellulose acetate trimellitate and carboxymethylcellulose sodium;acrylic acid polymers and copolymers, e.g., formed from acrylic acid,methacrylic acid, methyl acrylate, ethyl acrylate, methyl methacrylateand/or ethyl methacrylate, and other methacrylic resins that arecommercially available under the tradename Eudragit®. (Rohm Pharma;Westerstadt, Germany), including Eudragit®. L30D-55 and L100-55 (solubleat pH 5.5 and above), Eudragit®. L-100 (soluble at pH 6.0 and above),Eudragit®. S (soluble at pH 7.0 and above, as a result of a higherdegree of esterification), and Eudragits®. NE, RL and RS(water-insoluble polymers having different degrees of permeability andexpandability); vinyl polymers and copolymers such as polyvinylpyrrolidone, vinyl acetate, vinylacetate phthalate, vinylacetatecrotonic acid copolymer, and ethylene-vinyl acetate copolymer;enzymatically degradable polymers such as azo polymers, pectin,chitosan, amylose and guar gum; zein and shellac. Combinations ofdifferent coating materials may also be used. Multi-layer coatings usingdifferent polymers may also be applied.

The coating composition may include conventional additives, such asplasticizers, pigments, colorants, stabilizing agents, glidants, etc. Aplasticizer is normally present to reduce the fragility of the coating,and will generally represent about 10 wt. % to 50 wt. % relative to thedry weight of the polymer. Examples of typical plasticizers includepolyethylene glycol, propylene glycol, triacetin, dimethyl phthalate,diethyl phthalate, dibutyl phthalate, dibutyl sebacate, triethylcitrate, tributyl citrate, triethyl acetyl citrate, castor oil andacetylated monoglycerides. A stabilizing agent is used to stabilizeparticles in the dispersion. Typical stabilizing agents are nonionicemulsifiers such as sorbitan esters, polysorbates andpolyvinylpyrrolidone. Glidants are recommended to reduce stickingeffects during film formation and drying, and will generally representapproximately 25 wt. % to 100 wt. % of the polymer weight in the coatingsolution. One effective glidant is talc. Other glidants such asmagnesium stearate and glycerol monostearates may also be used. Pigmentssuch as titanium dioxide may also be used. Small quantities of ananti-foaming agent, such as a silicone (e.g., simethicone), may also beadded to the coating composition.

The particles can be prepared entirely from a therapeutic agent, or froma combination of the agent and a surfactant. The particles can be madeof a variety of materials. Both inorganic and organic materials can beused. For example, ceramics may be used. Polymeric and non-polymericmaterials, such as fatty acids, may be used to form aerodynamicallylight particles. Other suitable materials include, but are not limitedto, gelatin, polyethylene glycol, trehalose, and dextran. Particles withdegradation and release times ranging from seconds to months can bedesigned and fabricated, based on factors such as the particle material.

In addition to a therapeutic or diagnostic agent (or possibly otherdesired molecules for delivery), the particles can include excipientssuch as a sugar, such as lactose, a protein, such as albumin, and/or asurfactant.

Enteric Coated Capsules: “Gastric resistant natural polymer,” as usedherein, refers to natural polymers or mixtures of natural polymers whichare insoluble in the acidic pH of the stomach. “Film-forming naturalpolymer,” as used herein, refers to polymers useful for surface coatingsthat are applied by spraying, brushing, or various industrial processes,which undergo film formation. In most film-formation processes, a liquidcoating of relatively low viscosity is applied to a solid substrate andis cured to a solid, high-molecular-weight, polymer-based adherent filmpossessing the properties desired by the user. For most commonapplications, this film has a thickness ranging from 0.5 to 500micrometers (0.0005 to 0.5 millimeters, or 0.00002 to 0.02 inches).

“Gelling agent,” as used herein, refers to substances that undergo ahigh degree of cross-linking or association when hydrated and dispersedin the dispersing medium, or when dissolved in the dispersing medium.This cross-linking or association of the dispersed phase alters theviscosity of the dispersing medium. The movement of the dispersingmedium is restricted by the dispersed phase, and the viscosity isincreased.

Gastric resistant film-forming compositions containing (1) a gastricresistant natural polymer; (2) a film-forming natural polymer; andoptionally (3) a gelling agent, are described herein. Exemplary gastricresistant natural polymers include, but are not limited to, pectin andpectin-like polymers which typically consist mainly of galacturonic acidand galacturonic acid methyl ester units forming linear polysaccharidechains. Typically these polysaccharides are rich in galacturonic acid,rhamnose, arabinose and galactose, for example the polygalacturonans,rhamnogalacturonans and some arabinans, galactans and arabinogalactans.These are normally classified according to the degree of esterification.In high (methyl) ester (“HM”) pectin, a relatively high portion of thecarboxyl groups occur as methyl esters, and the remaining carboxylicacid groups are in the form of the free acid or as its ammonium,potassium, calcium or sodium salt. Useful properties may vary with thedegree of esterification and with the degree of polymerization. Pectin,in which less than 50% of the carboxyl acid units occur as the methylester, is normally referred to as low (methyl) ester or LM-pectin. Ingeneral, low ester pectin is obtained from high ester pectin bytreatment at mild acidic or alkaline conditions. Amidated pectin isobtained from high ester pectin when ammonia is used in the alkalinedeesterification process. In this type of pectin some of the remainingcarboxylic acid groups have been transformed into the acid amide. Theuseful properties of amidated pectin may vary with the proportion ofester and amide units and with the degree of polymerization. In oneembodiment, the gastric resistant natural polymer is pectin. The gastricresistant natural polymer is present in an amount less than about 5% byweight of the composition, e.g., from about 2 to about 4% by weight ofthe composition.

Exemplary film-forming natural polymers include, but are not limited to,gelatin and gelatin-like polymers. In an exemplary embodiment, thefilm-forming natural polymer is gelatin. A number of other gelatin-likepolymers are available commercially. The film-forming natural polymer ispresent in an amount from about 20 to about 40% by weight of thecomposition, e.g., from about 25 to about 40% by weight of thecomposition.

The compositions can optionally contain a gelling agent. Exemplarygelling agents include divalent cations such as Ca²⁺ and Mg²⁺. Sourcesof these ions include inorganic calcium and magnesium salts and calciumgelatin. The gelling agent is present in an amount less than about 2% byweight of the composition, e.g., less than about 1% by weight of thecomposition.

One or more plasticizers can be added to the composition to facilitatethe film-forming process. Suitable plasticizers include glycerin,sorbitol, sorbitans, maltitol, glycerol, polyethylene glycol,polyalcohols with 3 to 6 carbon atoms, citric acid, citric acid esters,triethyl citrate and combinations thereof. The concentration of the oneor more plasticizers is from about 8% to about 30% by weight of thecomposition. In one embodiment, the plasticizer is glycerin and/orsorbitol.

The film-forming composition can be used to prepare soft or hard shellgelatin capsules which can encapsulate a liquid or semi-solid fillmaterial or a solid tablet (Softlet®) containing an active agent and oneor more pharmaceutically acceptable excipients. Alternatively, thecomposition can be administered as a liquid with an active agentdissolved or dispersed in the composition.

The film-forming composition can be used to prepare soft or hardcapsules using techniques well known in the art. For example, softcapsules are typically produced using a rotary die encapsulationprocess. Fill formulations are fed into the encapsulation machine bygravity.

The capsule shell can contain one or more plasticizers selected from thegroup consisting of glycerin, sorbitol, sorbitans, maltitol, glycerol,polyethylene glycol, polyalcohols with 3 to 6 carbon atoms, citric acid,citric acid esters, triethyl citrate and combinations thereof.

In addition to the plasticizer(s), the capsule shell can include othersuitable shell additives such as opacifiers, colorants, humectants,preservatives, flavorings, and buffering salts and acids.

Opacifiers are used to opacify the capsule shell when the encapsulatedactive agents are light sensitive. Suitable opacifiers include titaniumdioxide, zinc oxide, calcium carbonate and combinations thereof.

Colorants can be used to for marketing and productidentification/differentiation purposes. Suitable colorants includesynthetic and natural dyes and combinations thereof.

Humectants can be used to suppress the water activity of the softgel.Suitable humectants include glycerin and sorbitol, which are oftencomponents of the plasticizer composition. Due to the low water activityof dried, properly stored softgels, the greatest risk frommicroorganisms comes from molds and yeasts. For this reason,preservatives can be incorporated into the capsule shell. Suitablepreservatives include alkyl esters of p-hydroxy benzoic acid such asmethyl, ethyl, propyl, butyl and heptyl (collectively known as“parabens”) or combinations thereof.

Flavorings can be used to mask unpleasant odors and tastes of fillformulations. Suitable flavorings include synthetic and naturalflavorings. The use of flavorings can be problematic due to the presenceof aldehydes which can cross-link gelatin. As a result, buffering saltsand acids can be used in conjunction with flavorings that containaldehydes in order to inhibit cross-linking of the gelatin.

Soft or hard capsules can be used to deliver a wide variety ofpharmaceutically active agents. Suitable agents include small molecules,proteins, nucleic acid, carbohydrates, lipids, and full organisms.

Fill formulations may be prepared using a pharmaceutically acceptablecarrier composed of materials that are considered safe and effective andmay be administered to an individual without causing undesirablebiological side effects or unwanted interactions. The carrier is allcomponents present in the pharmaceutical formulation other than theactive ingredient or ingredients. As generally used herein “carrier”includes, but is not limited to surfactants, humectants, plasticizers,crystallization inhibitors, wetting agents, bulk filling agents,solubilizers, bioavailability enhancers, pH adjusting agents, andcombinations thereof.

Alternatively, the composition can be administered as a liquid with anactive agent dissolved (e.g. solution) or dispersed (e.g., suspension)in the composition. Suitable active agents are described above. Thesolution or suspension may be prepared using one or morepharmaceutically acceptable excipients. Suitable excipients include, butare not limited to, surfactants, humectants, plasticizers,crystallization inhibitors, wetting agents, bulk filling agents,solubilizers, bioavailability enhancers, pH adjusting agents, flavorantsand combinations thereof.

Mucoadhesive Particles and methods of manufacturing: In general terms,adhesion of polymers to tissues may be achieved by (i) physical ormechanical bonds, (ii) primary or covalent chemical bonds, and/or (iii)secondary chemical bonds (e.g., ionic). Physical or mechanical bonds canresult from deposition and inclusion of the adhesive material in thecrevices of the mucus or the folds of the mucosa. Secondary chemicalbonds, contributing to bioadhesive properties, consist of dispersiveinteractions (e.g., Van der Waals interactions) and stronger specificinteractions, which include hydrogen bonds. The hydrophilic functionalgroups responsible for forming hydrogen bonds are the hydroxyl (—OH) andthe carboxylic groups (—COOH).

Adhesive polymeric microspheres have been selected on the basis of thephysical and chemical bonds formed as a function of chemical compositionand physical characteristics, such as surface area, as described indetail below. These microspheres are characterized by adhesive forces tomucosa of greater than 11 mN/cm². The size of these microspheres rangefrom between a nanoparticle to a millimeter in diameter. The adhesiveforce is a function of polymer composition, biological substrate,particle morphology, particle geometry (e.g., diameter) and surfacemodification.

Classes of Polymers Useful in Forming Bioadhesive Microspheres: Suitablepolymers that can be used to form bioadhesive microspheres includesoluble and insoluble, biodegradable and nonbiodegradable polymers.These can be hydrogels or thermoplastics, homopolymers, copolymers orblends, natural or synthetic. A key feature, however, is that thepolymer must produce a bioadhesive interaction between 110 N/m2 (11mN/cm2) and 100,000 N/m2 when applied to the mucosal surface of ratintestine.

In order for bioadhesive particles to embed themselves or becomeengulfed in the mucus lining the GI tract, the radius of the individualparticles should be as thick as the thickness of the natural mucouslayer. It has been shown that the gastric mucous layer thicknesstypically varies from 5 to 200.mu. in the rat and 10 to 400.mu. in man.Occasionally, however, it can reach thicknesses as great as 1000.mu. inman, as described by Spiro, R. G., “Glycoproteins,” Annual Review ofBiochemistry, 39, 599-638, 1970; Labat-Robert, J. & Decaeus, C.,“Glycoproteins du Mucus Gastrique: Structure, Fonction, et Pathologie,”Pathologie et Biologie (Paris), 24, 241, 1979; Allen, A., Hutton, D. A.,Pearson, J. P., & Sellers, L. A., “Mucus Glycoprotein Structure, GelFormation and Gastrointestinal Mucus Function” in Mucus and Mucosa, CibaFoundation Symposium 109 (eds. J. Nugent & M. O'Connor), pp. 137(London: Pitman, 1984). In the past, two classes of polymers haveappeared to show useful bioadhesive properties: hydrophilic polymers andhydrogels. In the large class of hydrophilic polymers, those containingcarboxylic groups (e.g., poly[acrylic acid]) exhibit the bestbioadhesive properties. One could infer that polymers with the highestconcentrations of carboxylic groups should be the materials of choicefor bioadhesion on soft tissues. In other studies, the most promisingpolymers were: sodium alginate, carboxymethylcellulose,hydroxymethylcellulose and methylcellulose. Some of these materials arewater-soluble, while others are hydrogels.

Rapidly bioerodible polymers such as poly[lactide-co-glycolide],polyanhydrides, and polyorthoesters, whose carboxylic groups are exposedon the external surface as their smooth surface erodes, are excellentcandidates for bioadhesive drug delivery systems. In addition, polymerscontaining labile bonds, such as polyanhydrides and polyesters, are wellknown for their hydrolytic reactivity. Their hydrolytic degradationrates can generally be altered by simple changes in the polymerbackbone.

Representative natural polymers include proteins, such as zein, modifiedzein, casein, gelatin, gluten, serum albumin, or collagen, andpolysaccharides, such as cellulose, dextrans, polyhyaluronic acid,polymers of acrylic and methacrylic esters and alginic acid.Representative synthetic polymers include polyphosphazines, poly(vinylalcohols), polyamides, polycarbonates, polyalkylenes, polyacrylamides,polyalkylene glycols, polyalkylene oxides, polyalkylene terephthalates,polyvinyl ethers, polyvinyl esters, polyvinyl halides,polyvinylpyrrolidone, polyglycolides, polysiloxanes, polyurethanes andcopolymers thereof. Synthetically modified natural polymers includealkyl celluloses, hydroxyalkyl celluloses, cellulose ethers, celluloseesters, and nitrocelluloses. Other polymers of interest include, but arenot limited to, methyl cellulose, ethyl cellulose, hydroxypropylcellulose, hydroxypropyl methyl cellulose, hydroxybutyl methylcellulose, cellulose acetate, cellulose propionate, cellulose acetatebutyrate, cellulose acetate phthalate, carboxymethyl cellulose,cellulose triacetate, cellulose sulfate sodium salt, poly(methylmethacrylate), poly(ethyl methacrylate), poly(butyl methacrylate),poly(isobutyl methacrylate), poly(hexyl methacrylate), poly(isodecylmethacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate),poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutylacrylate), poly(octadecyl acrylate)polyethylene, polypropylene,poly(ethylene glycol), poly(ethylene oxide), poly(ethyleneterephthalate), poly(vinyl acetate), polyvinyl chloride, polystyrene,polyvinyl pyrrolidone, and polyvinylphenol. Representative bioerodiblepolymers include polylactides, polyglycolides and copolymers thereof,poly(ethylene terephthalate), poly(butic acid), poly(valeric acid),poly(lactide-co-caprolactone), poly[lactide-co-glycolide],polyanhydrides, polyorthoesters, blends and copolymers thereof.

The pharmaceutically acceptable compounds of the invention will normallybe administered to a subject in a daily dosage regimen. For an adultsubject this may be, for example, an oral dose of at least one SCFA orcompound comprising a SCFA moiety of between 0.1 gram and 15 grams. Infurther embodiments, an oral dose of at least one SCFA or compoundcomprising a SCFA moiety can be between 0.5 gram and 10 grams. In stillfurther embodiments, an oral dose of at least one SCFA or compoundcomprising a SCFA moiety can be between 0.5 grams and 6 grams.

The pharmaceutical compositions may be administered 1, 2, 3, 4 or moretimes per day. Thus in particular embodiments, compositions formulated,for example, for topical administration may be administered multipletimes daily.

In one embodiment, compositions are contemplated comprising a 1:1 (w/w)ratio of a first and a second SCFA, wherein there may be 0.1, 0.2, 0.3,0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 grams ofa first SCFA. In other embodiments there may be a 2:1, 3:1, 4:1, 5:1;6:1, 7:1, 8:1, 9:1, or 10:1 (w/w) ratio of a first and a second SCFA,wherein there may be 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2,3, 4, 5, 6, 7, 8, 9, or 10 grams of a first SCFA. Of course, the ratioof a first and a second SCFA administered may be varied from thatdisclosed herein above. For example, any amount of a first SCFAincluding 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6,7, 8, 9, or 10 grams of a first SCFA may be administered with any amountof a second SCFA including 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9,1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 grams of a second SCFA. Such amounts ofeither supplement may be admixed in one composition or may be indistinct compositions.

Kits

The invention also includes a kit comprising compounds useful within themethods of the invention and an instructional material that describes,for instance, the method of administering compositions of the inventionas described elsewhere herein. In one embodiment, the kit comprises acomposition of the invention.

Experimental Examples

The invention is further described in detail by reference to thefollowing experimental examples. These examples are provided forpurposes of illustration only, and are not intended to be limitingunless otherwise specified. Thus, the invention should in no way beconstrued as being limited to the following examples, but rather shouldbe construed to encompass any and all variations which become evident asa result of the teaching provided herein.

Without further description, it is believed that one of ordinary skillin the art can, using the preceding description and the followingillustrative examples, make and utilize the present invention andpractice the claimed methods. The following working examples thereforeare not to be construed as limiting in any way the remainder of thedisclosure.

Example 1: Treatment of Skin Disorders with Short Chain Fatty Acids

Psoriasis is a chronic auto-inflammatory disease that causes raised,red, scaly patches to appear on the skin. It typically affects theoutside of the elbows, knees or scalp, though it can appear on anylocation. Some people report that psoriasis is itchy, burns and stings.Psoriasis is associated with other serious health conditions, such asdiabetes, heart disease and depression.

Current treatments of psoriasis include topical (creams, ointments, andphototherapy) and systemic (oral and injected medications) therapies.Topical creams and ointments include corticosteroids (overuse can causeskin thinning and resistance), retinoids, anthralin (normalizes skincell growth) and vitamin D analogs (treatment with retinoids, anthralinor vitamin D analogs can irritate skin), calcineurin inhibitors (disruptT-cell activation, continuous use is associated with increased risk ofskin cancer and lymphoma), as well as nonprescription moisturizers andsalicylic acid.

Phototherapy (UV light) includes the use of ultraviolet A (UVA) orultraviolet B (UVB) lights either alone or in combination withmedications. Short-term side effects include nausea, headache, burningand itching. Long-term side effects include dry and wrinkled skin,freckles, and increased risk of skin cancer, including melanoma.

Currently available systemic psoriasis therapies involve the use ofretinoids (side effects may include lip inflammation, hair loss, andsevere birth defects: women must avoid pregnancy for at least threeyears after taking the medication); chemotherapeutic drugs such asmethotrexate and cyclosporine (may cause a number of serious sideeffects, including liver damage, decreased production of red and whiteblood cells and platelets, increased risk of infections and cancer); andimmunomodulatory drugs or biologics (have strong effects on the immunesystem and may permit life-threatening infections). The side effects andhigh toxicity of the currently available treatments are undesirable formany patients suffering psoriasis.

SCFAs have never been used to treat any form of psoriasis. A majoradvantage of this approach is that there is little to no toxicityrelative to other currently available treatments. SCFAs were givenorally up to three times a day for as long as 3 weeks. The dose schedulewas one to two (1-2) 600 mg butyrate pills 3 times daily for 7 days to 4weeks followed by one (1) 600 mg butyrate pill 3 times daily for severalweeks to several months. Sodium, magnesium or calcium salts of butyratewere used for these treatments. When treatment was discontinued,psoriasis skin lesions reappeared. When treatments were started again,lesions disappeared once more. The immediate pre- and post-treatmentresults from 4 individuals taking butyrate are shown in FIG. 2 throughFIG. 5. Individuals with psoriasis taking butyrate capsules orally for3-4 weeks showed marked improvement or complete resolution of skinlesions.

For example, clinical trials can evaluate different combinations anddose schedules of selected SCFAs in healthy individuals (Phase 1) andamong patients with psoriasis (Phase II).

For example, the toxicity of single or combinations of SCFA is evaluatedin a dose escalation study to determine the level of toxicity and whattissues or organs are affected. Pharmacokinetic (PK) studies are to beconducted in mice to optimize the oral dose for further preclinicalwork. In the latter category, it will be important to see whether themolecules that are thought to mediate the pathogenesis of psoriasis inprior mouse studies are the same pathways inhibited in mice treated withSCFAs. For this work, two mouse models of psoriasis will be used. Thesepreclinical studies will also be validated by prior human studiesshowing that the pro-inflammatory molecules targeted by SCFAs in micewith psoriasis are the same ones that appear to mediate psoriasis inpeople. This work will determine whether oral administration of SCFAsresolves psoriasis lesions in mice. This appears to be the case inhumans taking butyrate orally. To optimize the dosage of a creamcontaining SCFAs, the cream will be applied to some but not otherlesions in mice with psoriasis. If only the treated lesions resolve,then the effect of SCFAs is local, but if application to some lesionsresults in resolution of all lesions, then the effect of SCFAs isprobably mediated by their systemic distribution. Depending upon theresults, parallel clinical trials will be performed in patients.

The materials and methods used in the experiments are now described.

Materials and Methods

SCFAs

SCFAs, consisting of sodium butyrate, magnesium butyrate, and calciumbutyrate were purchased commercially and manufactured by BodyBio.

Treatment with SCFAs

In all cases, no additional medication for psoriasis treatment was usedduring the butyrate regimen.

600 mg butyrate was administered orally 3 times daily to subjects havingmoderate to severe psoriasis for at least 2 weeks. 1200 mg butyrate wasadministered orally 3 times daily to a subject having severe psoriasisfor 3 weeks followed by administration of 600 mg butyrate orally 3 timesdaily for an extended period of time.

Lesions were evaluated before and after treatment.

The results of the experiments are now described.

FIG. 1 depicts a diagram showing signaling pathways that are affected bySCFAs. Of the proteins included on the diagram, IL-18, TLR3, IFN-γ,TNFα, TGF-β, MyD88, PI3K/Akt, JAK/STAT, Smad 2/3, Smad 4 and IL-10 werefound to be downregulated by SCFA treatment. The levels of IL-6, TRIF,PKR, TRAF2, TAK1 and TRAF6 were not evaluated for this study.

FIG. 2 shows results from a psoriasis patient undergoing a butyratetreatment regimen. The treatment regimen used was one (1) 600 mgbutyrate pill 3 times daily for 16 days. Images are of the patient'sleft elbow before and after treatment, the patient's right elbow beforeand after treatment, and the re-emergence of psoriasis on the patient'sleft elbow after 20 days the patient ceased the butyrate treatmentregimen.

FIG. 3 shows results from another psoriasis patient undergoing abutyrate treatment regimen. The treatment regimen used was two (2) 600mg butyrate pill 3 times daily for 10 days, then regimen was one (1)butyrate pill, three times/day for three weeks.

FIG. 4 shows results from a third psoriasis patient undergoing abutyrate treatment regimen. The treatment regimen used was two (2) 600mg butyrate pills 3 times daily for 7 days then one (1) 600 mg butyratepill 3 times daily for 24 days. Images are of the patient's left elbowbefore and after treatment. The patient consumed elevated doses ofalcohol during the treatment period.

FIG. 5 shows results from a fourth patient with severe psoriasis overabout 50% of the skin undergoing a butyrate treatment regimen. Patientstarted regimen as two (2) 600 mg butyrate pills 3 times daily for 4weeks followed by one (1) 600 mg butyrate pill 3 times daily for severalmonths. There was significant improvement, and no side effects werenoted during the treatment for long period of time. To confirm thatobserved positive effects were due to treatment with our formulation,patient was asked to stop regimen. Psoriasis came back in about 20-25days (pictures before). Patient started regimen as two (2) 600 mgbutyrate pills 3 times daily for 10 days then one (1) 600 mg butyratepills 3 times daily for 3 weeks and psoriasis disappeared (picturesafter). Images are of the patient's left hip before and after treatmentthe second treatment regimen, the patient's right hip before and afterthe second treatment regimen, and the patient's tailbone before andafter the second treatment regimen.

FIG. 6 shows results from a patient with plaque psoriasis. An initialtreatment regimen used was two (2) 600 mg butyrate pills 3 times dailyfor 20 days, then regimen was with a reduced dose, one (1) 600 mgbutyrate pills 3 times daily for 25 days. Significant improvement wasobserved 3-5 months after he stopped his treatment. Patient was nottaking any medicine before (at least 3 months), during and aftertreatment (at least 2 months).

Suggested Treatment:

Amount/capsule Daily dose Butyric acid 900 (or 1800) mg 3600 mgPropionic acid 100 (or 200) mg  400 mg Magnesium 10 (or 20) mg  40 mgVitamin D3 50 (or 100) IU  200 IU

Example 2: Combination Treatment of Skin Disorders Using Short ChainFatty Acids

This example is based, in part, on the discovery that administration ofat least one SCFA in combination with at least one second compound is aneffective approach for treating skin disease and disorders, includingpsoriasis. Without being bound by theory, it is expected that thecombination of at least one SCFA with at least one second compound willprovide an effective treatment for skin disorders. Additional compoundsthat are contemplated for use include a PDE4 inhibitor, ananti-inflammatory compound, a disease-modifying antirheumatic drug(DMARD), an immunosuppressant, a biologic agent, a Cox-2 inhibitor,apremilast or a combination thereof, and/or another agent. Non-limitingexamples of these and other compounds useful in the invention areprovided supra.

Suggested Treatment:

Combination treatment with apremilast: the daily oral dose is 1-2 g ofbutyrate, 100 mg propionate, 10-15 mg apremilast, 10-20 mg Magnesium,80-100 IU Vitamin D3, 50-100 IU Vitamin E (d-alpha-tocopherol acetate).

Palm and feet psoriasis is poorly responsive to treatment (these partsof the body are constantly “disturbed” from everyday activities). Theimpact of hand and foot psoriasis on patients' quality of life isextremely high (patients are unable to wear shoes comfortably or usetheir hands).

For such psoriasis, combination with topical ointments is important. Thedaily oral dose of SCFAs formulation should be used accompanied withapplying ointment consisting of:

⅖ part clobetazol (0.05%),

⅕ part calcipotriene (vit D, 0.005%),

⅕ part salicylic acid (10%),

⅕ part vit E (0.5%).

All these components (separately) are used as topical treatment.

Example 3: Use of Short Chain Fatty Acids in the Treatment of EyeDiseases and Disorders

Uveitis onset is often sudden, and the treatment and prognosis ofvarious uveitis entities vary greatly. Delayed treatment can lead toserious complications (detachment, blindness). It is critical first touse powerful drugs to stop disease progression. SCFAs can be used(orally and/or in the form of eyedrops) to sustain achieved therapeuticeffect (in particular, reduce inflammation). This will eliminatenegative side effects (including glaucoma, cataract) which are usuallyobserved after prolonged treatment with biologics and steroids (as incase of chronic or autoimmune uveitis). SCFAs can be also used in themixture with antibiotics (as eyedrops) and/or steroids (given at reduceddoses).

Clinical trials evaluate different combinations and dose schedules ofSCFA in healthy individuals (Phase 1) and among patients with uveitis(Phase II). Because some cases of uveitis are associated with abacterial or viral infection, the formulation also may containappropriate antibiotics or anti-viral compounds, respectively, alongwith the SCFAs.

Additional research includes testing single or combinations of SCFA in adose escalation study to evaluate toxicity, and what form that toxicitytakes (i.e., what tissues or organs are affected). Pharmacokinetic (PK)studies may be conducted in mice to understand what oral doses should beused for further preclinical work. It will be important to see whetherthe molecules that are thought to mediate the pathogenesis of uveitis inprior animal studies are the same pathways inhibited in animal modelstreated here with SCFAs.

For this work, two mouse models of uveitis are used. In one model,experimental autoimmune uveitis (EAU) is triggered by immunization withthe retinal antigens S-ag and IRBP coupled to Complete Freund's Adjuvant(CFA) and a B. pertussis toxin boost. The second model uses aIRBP161-180 peptide specific transgenic T cell receptor on a B10.RIIIbackground, which then develops spontaneous uveitis. These preclinicalstudies are also be validated by prior human studies showing that thepro-inflammatory molecules targeted by SCFAs in mice with uveitis arethe same molecules that appear to mediate uveitis in humans. This workconfirms that oral administration of SCFAs resolves uveitis lesions inmice.

Dry eye occurs when the quantity and/or quality of tears fail to keepthe surface of the eye sufficiently lubricated. Tears consist of oils,water, mucus, and more than 1500 different proteins that protect eyes.Vision in a dry eye disease may be affected because tears play animportant role in light focusing. Factors that can contribute to dry eyeinclude medications (antihistamines, antidepressants, birth controlpills, medications for Parkinson's disease, high blood pressure etc.),aging, rosacea, autoimmune disorders (Sjögren's syndrome, lupus,scleroderma, etc.), vitamin A deficiency, etc.

As a result, tears in an outer (lipid layer) produced by the Meibomianglands, a middle (aqueous layer) produced by the lacrimal glands, and aninner (mucin layer) produced by goblet cells became compromised.

The frequent use of artificial tears or other lubricating eye drops isthe main dry eye treatment. However, if dry eye is associated withinflammation, cyclosporine A, corticosteroids, tacrolimus, tetracyclinederivatives, etc. are used (such treatment demonstrated measurableclinical improvement). In such cases, butyrate treatment could beuseful.

Behçet's Disease (BD) is an autoimmune reaction that causes bloodvessels (including eye blood vessels) to become inflamed (BD of theEye). Inflammation inside of the eye occurs in more than half of thosewith BD and can cause blurred vision, pain, and redness. Other symptomsof the disease include blood clots and inflammation in the centralnervous system and digestive organs. Some people may become blind orseverely disabled. Corticosteroids and medications that suppress theimmune system may be prescribed to reduce inflammation.

Although the disease occurrence is mainly associated with a geneticfactor (human leukocyte antigen (HLA)-B51 antigen), molecular mechanismsinvolve increased neutrophil motility, elevated production of TNF-α andIL-17 and decreased production of IL-10 (all of which are regulated bySCFAs). It was shown that anti-TNF-α therapy suppresses effector T-celldifferentiation in BD patients with uveitis. Since butyrate suppressesTNF-α, treatment with SCFAs (orally) may provide protection frominflammation in BD.

Inflammation after cataract surgery can be persistent. Althoughcorticosteroids and non-steroidal anti-inflammatory drugs (NSAIDs) areused to treat inflammation (prophylactically or post-operatively), thereare no established guidelines for the treatment of inflammation inducedby cataract surgery. The long-term use of corticosteroids has raisedsafety concerns, especially with regard to elevated intraocularpressure. Surgical trauma triggers the arachidonic acid cascade (whichare converted to prostaglandins (PG) by activated COX-1 and COX-2enzymes), and PG are the most important inflammation mediators in thisdisease. Since butyrate was shown to inhibit COX-2 expression and PGproduction it can be used for treatment (in eyedrops).

Based on animal data, SCFA oral doses should be high to have atherapeutic effect.

Suggested Treatment:

The daily oral dose is 4-5 g of butyrate and 1.5-2 g of propionate inenteric coated, extended release capsules, twice/day.

Eyedrops containing 20-30 μM of both butyrate and 10-20 μM propionate(since blood plasma concentrations of these SCFAs are up to ˜30 μM) canbe also used.

In case of dry eyes, eyedrops should be given continuously, twice/day incombination with lubricants.

In other eye diseases, eyedrops should be given four times/day (in caseof disease flares), and twice/day up to few months, in combination withoral doses of SCFAs.

SCFAs can be used in the mixture with antibiotics and/or steroids.

For example, in case of infectious uveitis: during first 3-5 days, useeyedrops (or eye injection) with antibiotics/steroids (at the dosesprescribed by doctors), then during following 2-3 weeks, use eyedropscontaining a mixture of SCFAs (20-30 μM) and antibiotics/steroids (athalf of the prescribed doses), then continue eyedrops with SCFAs (20-30μM) only. Based on the type of disease and severity, oral SCFAs could begiven as well, alone or in combination with antibiotics. This couldaccelerate healing and minimize side effects.

Treatment of an Individual with Uveitis Using SCFA

Experiments were designed to treat a subject having infectious uveitis.In some instances, butyrate is administered to the subject. Aftercompletion of the butyrate therapy, the subject experiencessignificantly reduced inflammation.

Example 4: Use of Short Chain Fatty Acids in the Treatment of MacularDegeneration

Age related macular degeneration (AMD) causes damage to the macula (asmall area in the center of the retina). In the dry form of AMD, lipidaggregates (drusen) accumulate in the retina. The late stage of dry AMD(geographic atrophy) is characterized by degeneration of retinal pigmentepithelium cells and the overlying light-sensing retinal photoreceptors.“There is no way to treat dry AMD.” The Age-Related Eye Disease Studies(AREDS) found that daily intake of high-dose vitamins (vitamins E and C)and minerals (zinc oxide) can slow progression of the disease. In wetAMD, choroidal neovascularization occurs (newly immature blood vesselsgrow toward the outer retina from the underlying choroid). These bloodvessels leak fluids and cause scarring of the macula. The major factorthat contributes to vascularization is vascular endothelial growthfactor (VEGF). Most widely used anti-angiogenic FDA-approved therapies(eye injection) include pegaptanib, lucentis and VEGF-TRAP-Eye (AmbatiJ, Fowler B J. Mechanisms of age-related macular degeneration. Neuron,2012; 75(1):26-39), which improve or stabilize vision in the majority ofpatients. Butyrate was found to repress angiogenesis in vitro and invivo and reduce expression of pro-angiogenesis factors, includinghypoxia inducible factors (HIF-1a) and VEGF (Canani B, Di Costanzo M,Leone L. The epigenetic effects of butyrate: potential therapeuticimplications for clinical practice. Clinical Epigenetics, 2012; 4(1):4).Therefore, butyrate can be used for AMD treatment (eyedrops).

Clinical trials can evaluate different combinations and dose schedulesof selected SCFAs in healthy individuals (Phase 1) and then amongpatients with adult macular degeneration (AMD) (Phase II). Some studieshave shown a modest improvement in advanced AMD by eating aMediterranean diet, so some human trials may combine this diet with SCFAto look for additive or synergistic effects. However, human trials couldtake up to 2 or more years to see statistically significant results.

Additional research can include testing single or combinations of SCFAs(+/− dietary anti-oxidants) in a dose escalation study to look fortoxicity, and what form that toxicity takes (i.e., what tissues ororgans are affected). Pharmacokinetic (PK) studies can also be conductedin mice to get an idea of what oral doses should be used for furtherpreclinical work. In the latter category, it can be important to seewhether the molecules that are thought to mediate the pathogenesis ofmacular degeneration in prior animal studies are the same pathwaysinhibited in animal models treated here with SCFAs.

For this work, two mouse models of AMD can be used. In one model of dryAMD, a novel murine model of immune mediated retinal degeneration wasinduced by immunization with carboxyethylpyrrole (CEP)-modified albumin(CEP-MSA). CEP albumin adducts generated in the retinal photoreceptorsin response to oxidative stress. Patients with AMD also have circulatingCEP autoantibodies. In another model, CCL2/CX3CR1 KO mice, which aredeficient in macrophage/retinal microglia migration (and recruitment),develop degenerative changes characteristic of some AMD lesions within4-6 weeks of age. Here, the tissue response to oxidative stress is notfunctional (repair vs. continuing inflammation). Given that SCFAs blockoxidative stress, block angiogenesis (by down-regulating expression ofVEGF and PDGF), and down-regulate inflammatory responses, they may beeffective in preventing and/or resolving AMD. Such preclinical studiescan be validated by prior human studies showing that thepro-inflammatory molecules targeted by SCFAs in mice with AMD are thesame ones that are associated with AMD in people. This work can confirmthat oral administration of SCFAs resolves AMD lesions in mice.

Suggested Treatment:

The daily oral dose is 4-5 g of butyrate and 1.5-2 g of propionate inenteric coated, extended release capsules, twice/day.

Eyedrops containing 20-30 μM of both butyrate and 10-20 μM propionate(since blood plasma concentrations of these SCFAs are up to ˜30 μM) canbe also used. Eyedrops should be given four times/day (in case ofdisease flares), and twice/day up to few months, in combination withoral doses of SCFAs.

SCFAs can be used in the mixture with antibiotics and/or steroids.

Example 5: Treatment and Prevention of Autoimmune and Allergic Diseasesin C-Section Delivered Neonates with Short Chain Fatty Acids

Infants born by Cesarean section (C-section) experience an increasedrisk of type 1 diabetes (Cardwell et al., 2008, Diabetologia,51(5):726-35) and asthma (Thavagnanam et al., 2008, Clin Exp Allergy,38(4):629-33). These infants (born either at term or prematurely)acquire a gut microbiome which is characteristic of the maternal skin,and not the birth canal. This leaves the newborns susceptible to thedevelopment of allergic and autoimmune diseases and disorders, includingallergic rhinitis, gastroenteritis, inflammatory bowel disease, asthma,juvenile rheumatoid arthritis, food allergies, obesity, and type 1diabetes. The number of C-sections rose in the U.S. by about 50% between1996-2005, when it reached 30.2%, which is much higher than the optimallimit of 10-15% as recommended by WHO. Other countries with ratesgreater than 20% include Canada, the United Kingdom, Mexico, and Brazil,suggesting that increasing numbers of infants will be at risk forallergic and autoimmune diseases. The rate of C-sections for secondbirths, after a first child has been delivered by C-section, is about90%. Presently, there are no standard treatments for these and otherallergic/autoimmune diseases that may appear after C-section delivery.Some clinics try to partially restore the mother's missing microbes tobabies born via C-section by swabbing those infants with their mother'svaginal fluid within two minutes of birth. However, this practice iscontroversial. There is even evidence that this approach caninadvertently expose infants to maternal diseases.

Experiments were designed to assess the efficacy of using SCFAs toprevent, delay, or augment the onset of at least one autoimmune and/orallergic disease or disorder in a subject. As discussed herein, infantsdelivered by C-section experience an elevated risk of developingautoimmune and allergic diseases and disorders, and the number andfrequency of C-sections is increasing. Therefore, experiments weredesigned to deliver SCFAs to an expectant mother or a neonate deliveredby C-section to prevent at least one autoimmune and/or allergic diseaseor disorder disclosed herein.

The one or more SCFAs of the invention can be given orally up to threetimes a day for as long as two months. The dose schedule can beingestion of 6 tablets per day (600 mg/tablet) for one week, followed by3 tablets per day (600 mg/tablet) until the C-section is performed.Sodium, magnesium, or calcium salts of SCFAs can be used for thesetreatments. Alternatively, if a pregnant woman expecting a C-section isnot given SCFAs, the SCFAs could be mixed into food or drink provided tothe infant.

In some aspects, experiments were designed to have the SCFAs bedeveloped as supplements or nutraceuticals. Alternatively, the SCFAs canbe repackaged as enteric coated capsules that deliver time releasedproduct(s) for maternal use, or added as a supplement to baby formula orfood for at least the first 6-12 or 18-24 months of life. Clinicaltrials can evaluate different combinations and dose schedules ofselected SCFAs in healthy individuals (Phase I) and then among womengiving birth via C-section and to infants born to women via C-section(Phase II).

Additional experiments were designed to include testing single orcombinations of SCFAs in a dose escalation study to look for toxicity,and what form that toxicity takes (i.e., what tissues or organs areaffected). Pharmacokinetic (PK) studies can also be conducted in mice toget an idea of what oral doses should be used for further preclinicalwork. In the latter category, it is important to see whether themolecules that are thought to mediate the pathogenesis of theappropriate allergic or autoimmune diseases in prior mouse studies arethe same pathways inhibited in mice treated with SCFAs. For this work,appropriate mouse models can be used. These preclinical studies can alsobe validated by prior human studies showing that the pro-inflammatorymolecules targeted by SCFAs in the mouse models are the same ones thatappear to mediate the corresponding diseases in people. This work candetermine whether oral administration of SCFAs resolves the relevantdiseases or disorders (e.g., lesions) in mice. Parallel clinical trialscan be performed in women expecting C-section delivery during latepregnancy and among newborns that have arrived by C-section.

A major advantage of this approach is little to no toxicity relative toother currently available treatments, and at the same time, helping tore-establish gut homeostasis.

Infants born by C-section usually do not receive any interventions on aregular basis. However, Similac Pro-Advance and Similac Pro-Sensitivehave human milk oligosaccharides, which are prebiotic, like that foundin most breast milk. The present example contemplates a combination ofone or more SCFAs with one or more of these products.

Suggested Treatment:

For breastfed infants: the daily oral dose for healthy mothers is 1-2 gof butyrate and 0.5-1 g of propionate and acetate in enteric coated,extended release capsules, 3 times/day during first month, then half ofthat dose during next 3 months.

In the article (Xu J, Chen X, Yu S, et al. Effects of Early Interventionwith Sodium Butyrate (SB) on Gut Microbiota and the Expression ofInflammatory Cytokines in Neonatal Piglets. PLoS One, 2016;11(9):e0162461), early intervention with sodium butyrate modulated theileum inflammatory cytokines in neonatal piglets with low impact onintestinal microbial structure, which suggests oral administration of SBmay have a benefit role in the health of neonatal piglets (1 and 7 daysold). The oral dose was ˜10 ml of 150 mmol/L (piglets consumed 10 ml of˜150 mmol/L of sodium butyrate which is equivalent to ˜165 mg). Forsupplementation in formula, a suggested treatment regimen is usingescalation doses: start with low concentrations (daily dose is 80-100 mgof butyrate, 20-30 mg of acetate and propionate during first month),then increase dose to 100-120 mg of butyrate and 30-40 mg of acetate andpropionate during next 3-5 months).

Example 6: Use of Short Chain Fatty Acids in the Treatment of Vasculitis

Vasculitis is a rare autoimmune disease which involves inflammation ofthe blood vessels, arteries, veins or capillaries and can affect peopleof all ages. Vasculitis also may be linked to certain blood cancers(leukemia and lymphoma). Different types of vasculitis are classifiedaccording to the size and location of the blood vessels that areaffected. Common vasculitis treatment includes corticosteroids andcytotoxic drugs. Some molecular mechanisms of the pathogenesis ofvasculitis are outlined in our patent application. GPR-109a pathway wasalso shown to play an important role (Chai J T, Digby J E, Choudhury RP. GPR109A and vascular inflammation. Curr. Atheroscler. Rep. 2013;15(5):325). Activation of GPR-109a receptor down-regulates NF-κB, and anumber of selective GPR109A agonists have been developed by Merck, GSKand other companies. This receptor is activated by butyrate (at EC50˜1.5mM) and niacin.

Clinical trials will evaluate different combinations and dose schedulesof selected SCFAs in healthy individuals (Phase I) and then amongpatients with vasculitis (Phase II). At this point, cyclophosphamide orglucocorticoids are used most often for treatment, althoughmethotrexate, azathioprine and mycophenolate have also been used. Theproblem is that these are highly toxic therapeutic approaches that limittheir application in patients. More specific biologic therapies usingmonoclonal antibodies against TNFβ, IL-1 or IL-6 require evaluation.Given that the inflammatory processes in atherogenesis, most forms ofvasculitis, and aneurysms share many characteristics, it is likely thatintervention with SCFAs will show broader application in re-establishinghomeostasis among these various disease states.

Additional research will include testing single or combinations of SCFAsin a dose escalation study to look for toxicity, and what form thattoxicity takes (i.e., what tissues or organs are affected).Pharmacokinetic (PK) studies will also have to be conducted in mice toget an idea of what oral doses should be used for further preclinicalwork. In the latter category, it will be important to see whether thetargets that are thought to mediate the pathogenesis of vasculitis inanimal studies are the same targets/pathways inhibited in animal modelstreated here with SCFAs.

For this work, two mouse models of ANCA associated vasculitis can beused, since these are fairly well characterized. In one model, MPOknockout mice (MPO-/-mice) are immunized with MPO. These animals arethen irradiated and reconstituted with syngeneic, wild type bone marrow.Since antibody producing plasma cells are relatively radio-resistant,circulating anti-MPO levels are maintained. The bone marrowreconstitutes neutrophils which are then available to bind anti-MPO.Mice then develop crescentic glomerulonephritis and urine abnormalitiesby 8 weeks post bone marrow transplant. Without being bound by anyparticular theory, given that neutrophils expressing MPO is also atarget in human vasculitis, and that SCFAs down-regulate MPO expressionby blocking NF-κB activity in macrophages (thereby blocking thesecretion of factors by macrophages that activate neutrophils), it isproposed that the administration of SCFAs can block the development ofvasculitis in this animal model.

Another model involves immunization of wild type C57BL/6 mice with humanor mouse MPO, leading to the induction of both humoral and cellularresponses to MPO. While this alone does not result in disease, thepassive transfer of polyclonal glomerular binding antibody results inthe accumulation of neutrophils into the glomeruli. The MPO in theseneutrophils is then targeted by antigen specific CD4+ T cells, resultingin autoimmune glomerulonephritis, which is seen in about 50% of patientswith ANCA vasculitis. Other animal models are available if needed.Without being bound by any particular theory, given that SCFAs promotethe differentiation of T cells to the Treg phenotype, it is hypothesizedthat the administration of SCFAs can block or ameliorate the developmentof autoimmune glomerulonephritis.

Such preclinical studies can be validated by prior human studies in theliterature showing that the pro-inflammatory molecules targeted by SCFAsin mice with vasculitis are the same ones that are associated withvasculitis in people. This work can determine whether oraladministration of SCFAs resolves vasculitis in mice.

The models outlined in this Example may not cover all aspects ofvasculitis pathogenesis, and for many forms of vasculitis, there is noanimal model available. For example, regarding ANCA vasculitis, there isno reproducible model of granulomatous PR3-ANCA. However, the modelsused can establish proof-of-principle as to whether SCFAs should betaken into human clinical trials against vasculitis.

Suggested Treatment:

The daily oral dose is 5-6 g of butyrate in enteric coated, extendedrelease capsules, 3 times/day, during first month, then 3-4 g ofbutyrate, twice a day, during several months. High doses should be usedin case of flares.

Butyrate could be used in combination with reduced doses of steroids andchemodrugs.

Example 7: Use of Short Chain Fatty Acids to Treat Selected Lymphomas

Since the present example is an idea based upon the properties of SCFAs,combined with the difficulties achieving long term remission andpreventing relapse of selected lymphomas, it can be useful to test thisapproach in 1-2 preclinical models for each selected lymphoma toestablish proof-of-principle for human trials. The invention itselfsimply involves oral administration of SCFAs at the time of diagnosiseither alone or in combination with standard of care therapy. Moreover,in mouse models, treatment prior to the development of lymphoma canestablish whether SCFAs can delay or block tumor development. Given thatthese cancers in people are often accompanied by multiple geneticaberrations that may be present prior to the clinical phase of thedisease, and that some of these tumors may have associated hereditarypredisposition, suggest that SCFAs may (at a minimum) help to preventrecurrence/relapse.

SCFAs could be used (1) alone or in combination with standard of caretreatments for selected lymphomas that depend upon constitutiveactivation of NF-κB, since the SCFAs used herein block NF-κB activity,(2) to delay or prevent lymphoma recurrence/relapse, (3) to decrease thetoxicity profiles of standard of care treatments, thereby increasinglong term quality of life, (4) to possibly intervene in high riskpatients prior to the appearance of lymphoma, and (5) to targetlymphomas by virtue of SCFAs histone deacetylase (HDAC) inhibitoryactivity, which is not part of any other therapeutic approach inpractice or in development. These features will help to extend theinvention to better treat patients with high tumor loads.

Suggested Treatment:

The daily oral dose is 4-5 g of butyrate in enteric coated, extendedrelease capsules, 3 times/day continuously.

Treatment with butyrate could be done in combination with reduced dosesof steroids and chemodrugs.

Example 8: Use of Short Chain Fatty Acids to Treat Leukemias

Philadelphia chromosome negative (Ph−) myeloproliferative neoplasms(MPNs) include polycythemia vera (PV), essential thrombocytosis (ET),and myelofibrosis (MF), all of which may evolve into acute myeloidleukemia (AML). MPNs are characterized by constitutive activation of themyeloproliferative leukemia (MPL) oncogene and the signaling molecule,JAK2 (usually by mutation). STAT-1, -3 and -5, MAPK, ERK and AKT/PI3K,all support cytokine independent growth in PV, ET and MF. The tumorsuppressor and epigenetic modifier, ten-eleven translocation 2 (TET2) isalso frequently mutated in these cells, suggesting that activation ofJAK2 and inactivation of TET2 are drivers of AML. They blockdifferentiation and promote self-renewal. Ph-MPNs are relatively rare,ranging from 90-120 cases per 100,000. A variety of tyrosine kinaseinhibitors (TKIs) have been developed to block JAK/STAThyper-activation, but this is not curative in many patients and isassociated with significant toxicity.

Butyrate has been shown to block nuclear translocation of STAT1, whichblocks JAK2 activation. Further, butyrate and propionate are HDACinhibitors. Elevated HDAC activity inhibits retinoic acid (RA) induceddifferentiation, so it is expected that butyrate/propionate+retinoicacid may prevent the progression and/or relapse of MPNs.

Pediatric ALL is also characterized by constitutive activation of theJAK/STAT pathway, which is inhibited by butyrate. In addition, butyrateactivates p53, which is otherwise down-regulated in ALL. Further, theSCFA valproate inhibits TGFbeta1 and PI3K signaling in ALL, all of whichsuggest that SCFA intervention would demonstrate efficacy in thiscontext. This is especially important in pediatric and adult ALL, sinceminimal residual disease is common after initial chemotherapy andrelapse time is characteristically short.

Since the present example is based upon the properties of SCFAs,combined with the difficulties achieving long term remission andpreventing relapse of AML and other leukemias, it would be useful totest this approach in 1-2 preclinical models for each selected leukemiato establish proof-of-principle for human trials. The invention itselfincludes oral administration of one or more SCFA therapeutic compoundsat the time of diagnosis, either alone or in combination with standardof care therapy. Moreover, in mouse models, treatment initiated at thetime of diagnosis will establish whether SCFAs can delay or block tumorprogression. Given that these cancers in people are often accompanied bymultiple genetic aberrations that may be present prior to the clinicalphase of the disease, and that some of these tumors may have associatedhereditary predisposition, suggest that SCFAs may (at a minimum) help toprevent recurrence/relapse.

Experiments include introducing AML or ALL cell lines (or banked primarycells from patients with the same disease) into immunodeficient mice(NOD/SCID) followed by SCFA treatment or placebo for different periodsof time. Periodic blood samples will be analyzed for minimal residualdisease by PCR amplification of the genetic rearrangements that arecommon in AML or ALL in clinical samples or known to be associated witha particular cell line that was used for injection. If commerciallyavailable transgenic mice carrying the major genetic lesionscharacteristic of AML or ALL are commercially available, thenexperiments could be planned to test whether SCFAs could alter diseasepathogenesis.

SCFAs could be used (1) alone or in combination with standard of caretreatments for selected leukemias (AML and ALL) that depend uponconstitutive activation of JAK/STAT, since the SCFAs used herein blockJAK/STAT activity, (2) to delay or prevent leukemia recurrence/relapse,(3) to decrease the toxicity profiles of standard of care treatmentsstarted at the time of diagnosis, thereby increasing long term qualityof life. These features will help to extend the invention to bettertreat patients with high tumor loads.

Currently, treatment includes two phases. The first phase, referred toas induction, often includes chemotherapy with ara-C and daunomycin,which lowers the white blood cell count and destroys the bone marrow.Sometimes a third drug, cladribine (Leustatin, 2-CdA), is given as well.Induction is considered successful if remission is achieved. However,induction leaves patients susceptible to fatal infections for up toseveral weeks, and during that time, they remain hospitalized. Thesecond phase, referred to as consolidation, which includes long termhigh dose ara-C. Alternatively, allogeneic or autologous stem celltransplant could be considered, although consolidation is associatedwith increased risk of death.

Constitutively activated HDAC has been observed in many types ofleukemias, and so synthetic HDAC inhibitors have been proposed asinterventional strategies. However, none have entered human clinicaltrials, although valproate and phenylbutyrate are under evaluation forselected lymphomas. In acute promyelocytic leukemia (APL), a variant ofAML, inhibition of HDAC1/2 promotes differentiation and apoptosis.However, treatment of preleukemic cells with the same HDAC inhibitorspromote cell proliferation, suggesting that SCFAs may be of use to blocktumor relapse, but may not prevent tumor onset. The potential advantageof using the mixture of SCFAs including butyrate and propionate for AMLrelapse is that they will be efficacious without appreciable sideeffects. Since SCFA associated HDAC inhibitory activity is reversible,the present example provides a potentially sound way of balancingefficacy while lowering the risk of toxic or other adverse effectscommon in other therapeutic approaches. This may be especially true inthe context of dealing with tumor recurrence/relapse where combinationtherapies may turn out to be the best option.

Suggested Treatment:

The daily oral dose is 4-5 g of butyrate in enteric coated, extendedrelease capsules, 3 times/day continuously.

Treatment with butyrate could be done in combination with reduced dosesof steroids and chemodrugs.

Example 9: Use of Short Chain Fatty Acids in Treatment or Prevention ofAdverse Effects Associated with CAR-T Therapy

The probability of the present example working in preclinical models ishigh given the goal of cancer immunotherapy and the knownimmunomodulatory functions of SCFAs. SCFAs can be taken orally, arenon-toxic, and have no problem crossing both the gut epithelium andblood-brain barrier, which are combined advantages not available byother approaches. Since immune-modulation by SCFAs is reversible, thepresent example provides a sound way of balancing efficacy whilelowering the risk of adverse effects for CAR-T and other cancerimmunotherapeutic approaches.

For example, in one aspect, the invention provides an adjunct therapy to(1) ameliorate a cytokine storm seen in some patients given CAR-T, (2)provide neuroprotection for patients experiencing these adverse effects,(3) immune-modulate CAR-T cell activity without using cytotoxicchemotherapy or systemic corticosteroids, (4) also have anti-tumorproperties by virtue of SCFAs histone deacetylase (HDAC) inhibitoryactivity in both liquid and solid tumors where the Warburg effect shuntsSCFAs to the nucleus, and (5) SCFAs are broadly anti-inflammatorybecause they modulate NF-κB, which directly stimulates production ofIL-6, TNFα, and IFNγ, all of which are commonly elevated in a cytokinerelease syndrome (CRS). SCFAs also down-regulate the production of TNFαand IL-6 from macrophages, thereby potentially ameliorating macrophageactivation syndrome, which is also a documented adverse effect of CAR-Ttherapy. This will help to extend the invention to better treat patientswith high tumor loads.

Two CD-19 targeted CAR-T therapies are approved by FDA for the treatmentof children with acute lymphoblastic leukemia and for adults withadvanced lymphomas.

Cytokine Release Storm (CRS) is considered to be an “on-target’ sideeffect, meaning T-cells are active. However, it can lead to dangerouslyhigh fever and quick drop in blood pressure. Patients experiencingsevere CRS all had particularly high levels of cytokine IL-6 (secretedby T cells and macrophages in response to inflammation). Tocilizumab(which blocks IL-6 activity) become a standard therapy for managing CRS.The approach worked, resolving the problem in most patients. However,patients treated with Tocilizumab are at very high risk for developingserious infections that may lead to hospitalization or death.

Butyrate (Yuan H, Liddle F J, Mahaj an S, Frank D A. IL-6-inducedsurvival of colorectal carcinoma cells is inhibited by butyrate throughdown-regulation of the IL-6 receptor. Carcinogenesis. 2004;25(11):2247-55) and propionate (Nastasi C, Fredholm S, Willerslev-OlsenA, et al. Butyrate and propionate inhibit antigen-specific CD8(+) T cellactivation by suppressing IL-12 production by antigen-presenting cells.Sci Rep. 2017; 7(1):14516) were shown to inhibit IL-6, thus can beconsidered as a co-treatment option.

SCFAs should be given in parallel to chemotherapy: several days beforeCAR-T cell infusion (only SCFAs), and up to several weeks after CAR-Tcell infusion (SCFAs in combination with significantly reduced doses ofchemodrug(s)). The daily oral dose is 5-6 g of butyrate and 2-3 g ofpropionate in enteric coated, extended release capsules, 3 times/day.

The disclosures of each and every patent, patent application, andpublication cited herein are hereby incorporated herein by reference intheir entirety. While this invention has been disclosed with referenceto specific embodiments, it is apparent that other embodiments andvariations of this invention may be devised by others skilled in the artwithout departing from the true spirit and scope of the invention. Theappended claims are intended to be construed to include all suchembodiments and equivalent variations.

What is claimed is:
 1. A composition comprising at least one compoundselected from the group consisting of: a short chain fatty acid (SCFA),a SCFA precursor, a SCFA biosynthesis precursor, a compound comprising aSFCA moiety, a derivative thereof, and a combination thereof.
 2. Thecomposition of claim 1, wherein the composition comprises at least oneSCFA moiety linked to at least one additional moiety.
 3. The compositionof claim 2, wherein the at least one additional moiety comprisespolyethylene glycol (PEG).
 4. The composition of claim 3, wherein thecomposition hydrolyzes under a low pH condition to yield PEG and a SCFA.5. The composition of claim 1, wherein the composition comprises atleast one compound selected from the group consisting of: a) at leastone SCFA or SCFA moiety selected from the group consisting of: aceticacid, butyric acid (BA), C3-C12 fatty acids, C3-C10 fatty acids, C3-C8fatty acids, methoxyacetic acid, valproic acid (VPA), propionic acid,3-methoxypropionic acid, ethoxyacetic acid, formic acid, isobutyricacid, tributyrin, butyrate, propionate, N-acetylbutyrate (as well asother forms of butyrate, e.g., phenylbutyrate, isobutyrate,pivaloyloxymethyl butyrate, monoacetone glucose 3-butyrate), isovalericacid, valeric acid, isocaproic acid, caproic acid, lactic acid, succinicacid, pyruvic acid, octanoic acid, dodecanoic acid,(4R)-4-hydroxypentanoic acid, 2-ethylhydracrylic acid,2-hydroxy-3-methylpentanoate, 2-hydroxy-3-methylpentanoic acid,2-methylbut-2-enoic acid, 2-oxobutanoic acid, 3-hydroxypentanoic acid,3-methylbut-2-enoic acid, butenoic acid, methylbutyric acid,dimethylbutyric acid, pentadienoic acid, pentenoic acid, pivalic acid,propynoic acid and a combination thereof; b) at least one SCFA precursoror SCFA precursor moiety selected from the group consisting of: a saltof lactate, a salt of succinate, a salt of formate, 1,2-propenedol,trypamine, indole, indole-3-acetate, and a combination thereof; and c)at least one SCFA biosynthesis precursor or SCFA biosynthesis precursormoiety selected from the group consisting of: an acetyl-CoA carboxylaseinhibitor, an adenosine monophosphate kinase (AMPK) activator, vitaminD, and a combination thereof.
 6. The composition of claim 1, wherein thecomposition comprises at least one SCFA selected from the groupconsisting of: butyrate, propionate and acetate.
 7. A pharmaceuticalcomposition comprising a composition of claim
 1. 8. The composition ofclaim 7, wherein the composition comprises 100 milligrams (mg) to 6grams (g) of at least one SCFA.
 9. The composition of claim 8, whereinthe composition is an enteric coated, extended release capsule.
 10. Thecomposition of claim 9, wherein the composition comprises 600 mgbutyrate.
 11. The composition of claim 9, wherein the compositioncomprises 900 mg butyrate, 100 mg propionate, 10 mg apremilast, 10 mgmagnesium, and 50 IU vitamin D3.
 12. The composition of claim 7, whereinthe composition is formulated as an eyedrop.
 13. The composition ofclaim 11, wherein the composition comprises at least one selected fromthe group consisting of: a) 10 micromolar (μM) to 100 μM butyrate; b) 10μM to 100 μM propionate; and c) 10 μM to 100 μM butyrate and 10 to 100μM propionate.
 14. A method for treating or preventing a disease ordisorder, comprising administering to a subject a composition of claim7.
 15. The method of claim 14, wherein the disease or disorder comprisesat least one selected from the group consisting of: a skin disease ordisorder, an allergic or autoimmune disease or disorder, an eye diseaseor disorder, an adverse effect associated with immunotherapy, cancer anda combination thereof.
 16. The method of claim 14, wherein the methodcomprises administering a composition comprising 100 mg to 6 g butyrate.17. The method of claim 16, wherein the method comprises administeringthe composition at least once daily for at least one week.
 18. Themethod of claim 14, further comprising administering at least oneadditional treatment or therapeutic agent to the subject.
 19. The methodof claim 18, wherein the at least one additional treatment isimmunotherapy.
 20. The method of claim 14, wherein the disease is a skindisease or disorder and further wherein the method comprises: a)administering a dose of at least 900 mg of butyrate and at least 100 mgpropionate in enteric coated, extended release capsules, 3 times/day forat least 2 weeks.
 21. The method of claim 20, wherein the method furthercomprises: b) administering a daily oral dose of at least 900 mg ofbutyrate, twice a day for at least 2 weeks, wherein step b) follows thecompletion of the at least 2 weeks of step a).
 22. The method of claim14, wherein the disease is a skin disease or disorder and furtherwherein the method comprises: administering a daily oral dose of atleast 900 mg of butyrate, at least 100 mg propionate, and at least 10 mgapremilast in enteric coated, extended release capsules, 3 times/day forat least 2 weeks.
 23. The method of claim 14, wherein the disease is aneye disease or disorder and further wherein the method comprises:administering a daily oral dose of at least 4 g of butyrate and at least1.5 g propionate in enteric coated, extended release capsules, 3times/day for at least 2 weeks.
 24. The method of claim 14, wherein thedisease is an eye disease or disorder and further wherein the methodcomprises: administering eyedrops containing at least 20 μM of butyrateand at least 10 μM propionate at least 2 times/day for at least 3 days.25. The method of claim 14, wherein the disease is an adverse effectassociated with immunotherapy and further wherein the method comprises:administering a daily oral dose of at least 5 g of butyrate and at least2 g propionate in enteric coated, extended release capsules, 3 times/dayfor at least 2 weeks.
 26. The method of claim 25, wherein theadministering is performed at a time selected from the group consistingof: prior to an immunotherapy, concurrent with an immunotherapy,subsequent to an immunotherapy, and a combination thereof.
 27. Themethod of claim 14, wherein the disease is selected from the groupconsisting of an allergic disease, an autoimmune disease and asthma, andfurther wherein the method comprises: administering a daily oral dose ofat least 1 of butyrate, at least 0.5 g propionate, and at least 0.5 gacetate in enteric coated, extended release capsules, 3 times/day for atleast 2 weeks.
 28. The method of claim 14, wherein the disease isselected from the group consisting of an allergic disease, an autoimmunedisease and asthma in a C-section delivered neonate, and further whereinthe method comprises: administering to the neonate a daily oral dose ofat least 80 mg of butyrate, at least 20 mg propionate, and at least 20mg acetate as a nutraceutical.
 29. The method of claim 14, wherein thedisease is vasculitis and further wherein the method comprises: a)administering a daily oral dose of at least 5 g of butyrate in entericcoated, extended release capsules, 3 times/day for at least 2 weeks. 30.The method of claim 29, wherein the method further comprises: b)administering a daily oral dose of at least 3 g of butyrate, twice a dayfor at least 2 weeks, wherein step b) follows the completion of the atleast 2 weeks of step a).